TW201908734A - Fluid control - Google Patents

Abstract

The present invention relates to a microfluidic assay system and associated reading device, as well as the individual components themselves. The present invention also relates to methods of conducting assays, using a disposable system and associated reading device, as well as kits for conducting assays.

Description

流體控制  Fluid control  

本發明係關於微流控檢定系統及相關聯讀數裝置，以及個別部件本身。本發明亦係關於使用拋棄式系統及相關聯讀數裝置來進行檢定之方法，以及進行檢定之套組。 The present invention relates to microfluidic assay systems and associated reading devices, as well as to individual components themselves. The invention also relates to methods for performing assays using disposable systems and associated reading devices, as well as kits for performing assays.

微流控盒常規用於執行各種檢定，亦即生物及化學及/或生理化學檢定，並且檢定之結果經常使用將該盒引入其中的相關聯讀數器裝置來判定。 Microfluidic cartridges are routinely used to perform various assays, i.e., biological and chemical and/or physicochemical assays, and the results of the assay are often determined using associated reader devices into which the cartridge is introduced.

通常需要在盒內之流體運動以便確保樣本能夠接觸試劑，該等試劑置放在盒中並且能夠與可存在於樣本中之一或多個目標分析物反應。在與一或多個試劑反應之後，通常需要將樣本從已經發生反應之區域移除，以使得可發生進一步反應，或僅允許偵測任何反應產物，此等目標可能在樣本保留在原位時，由於例如光干涉而導致難以達成。 Fluid movement within the cartridge is typically required to ensure that the sample is capable of contacting the reagents, which are placed in the cartridge and are capable of reacting with one or more analytes of interest that may be present in the sample. After reacting with one or more reagents, it is often necessary to remove the sample from the area where the reaction has occurred so that further reactions may occur, or only allow any reaction product to be detected, such targets may be retained while the sample remains in place It is difficult to achieve due to, for example, light interference.

盒內之流體運動可單獨藉由毛細管作用來發生，或盒內之流體之控制可藉由主動力或經由機構來實現，該主動力例如藉由使用可存在於盒中之微泵及閥來提供，該等機構存在於讀數器裝置中，該等機構被設計來與盒相互作用並且將流體泵送至盒中及/或將流體從盒中泵送出以便控制盒本身內之流體運動，參見例如EP2613881。或者可使用毛細管作用與主 動力之組合。 Fluid movement within the cartridge can occur solely by capillary action, or control of the fluid within the cartridge can be achieved by primary power or via a mechanism, such as by using a micropump and valve that can be present in the cartridge. Provided, the mechanisms are present in a reader device that is designed to interact with the cartridge and pump fluid into the cartridge and/or pump fluid out of the cartridge to control fluid movement within the cartridge itself, See, for example, EP2613881. Alternatively, a combination of capillary action and primary power can be used.

舉例而言，US7238324描述使用毛細管作用及應用外部泵的微流控裝置。允許樣本藉由毛細管作用、經由第一埠進入微流控盒並且流至感測腔室，其中發生檢定。在檢定反應之後，藉由使用外部泵、經由第二埠，將液體引入晶片中。此液體之用途係洗掉原始樣本，僅留下可被偵測的反應產物。然而，此意味著必須將單獨液體從外部並且經由另一個埠來引入晶片，該埠可被堵住及/或易受污染。此外，隨著時間的推移，液體可被污染或降解。因此，需要提供微流控盒，其中不需要從盒外部引入除了樣本以外的流體及/或具有除了引入樣本之樣本埠以外的埠。已知一些設計在盒本身內包括液體填充囊以便提供合適清洗試劑/緩衝液等，但是此舉顯著增加盒之複雜性及費用並且液體試劑亦可易發生降解。 For example, US Pat. No. 7,238,324 describes a microfluidic device that uses capillary action and applies an external pump. The sample is allowed to enter the microfluidic cartridge via the first enthalpy by capillary action and flow to the sensing chamber where the assay occurs. After the assay is reacted, liquid is introduced into the wafer via an external pump using a second pump. The use of this liquid is to wash away the original sample, leaving only the reaction product that can be detected. However, this means that a separate liquid must be introduced into the wafer from the outside and via another crucible, which can be blocked and/or susceptible to contamination. In addition, the liquid can be contaminated or degraded over time. Therefore, there is a need to provide a microfluidic cartridge in which it is not necessary to introduce a fluid other than the sample from the outside of the cartridge and/or have a crucible other than the sample of the introduced sample. Some designs are known to include a liquid-filled bladder within the cartridge itself to provide a suitable cleaning reagent/buffer, etc., but this significantly increases the complexity and expense of the cartridge and the liquid reagent can also be susceptible to degradation.

US5821399描述一種系統及盒，其使用導電率來量測樣本及樣本之間的流體。在盒內之流體填充囊中提供沖洗流體，其可在盒內傳送並且根據其導電率藉由讀數器來偵測。可容易地判定樣本與沖洗流體或空氣部分之間之導電率之差異。 No. 5,821,399 describes a system and cartridge that uses electrical conductivity to measure fluid between a sample and a sample. A flushing fluid is provided in the fluid-filled bladder within the cartridge that can be transported within the cartridge and detected by the reader based on its conductivity. The difference in conductivity between the sample and the flushing fluid or air portion can be readily determined.

WO2013154946描述微流控系統，其使用毛細管作用與氣體壓力之組合來控制微流控裝置內之液體樣本之運動。最初，引入裝置中之液體樣本藉由毛細管作用部分地沿著毛細管通道來傳送。當液體前移時，遠端氣體-液體界面處之氣體壓力增加一定量，該增加量足以停止液體的運動。為了引發液體樣本之進一步運動，連接至毛細管通道之遠端部分的泵將作用於液體樣本之遠端氣體-液體界面之氣體的壓力減少一定量，該減少量足以允許液體樣本藉由毛細管作用進一步沿著微流控裝置之毛細管通道 移動。 WO2013154946 describes a microfluidic system that uses a combination of capillary action and gas pressure to control the movement of a liquid sample within a microfluidic device. Initially, the liquid sample introduced into the device is transported partially along the capillary channel by capillary action. When the liquid is advanced, the gas pressure at the distal gas-liquid interface is increased by an amount sufficient to stop the movement of the liquid. In order to initiate further movement of the liquid sample, the pump connected to the distal portion of the capillary channel reduces the pressure of the gas acting on the distal gas-liquid interface of the liquid sample by an amount sufficient to allow the liquid sample to further act by capillary action Move along the capillary channel of the microfluidic device.

US5096669描述結合讀數器裝置來使用的拋棄式裝置。樣本可最初經由毛細管作用來抽吸至拋棄式裝置中並且在該裝置內之樣本之進一步運動可藉由讀數器自動地按壓該裝置內的包含可撓性膜的氣囊來實現，以導致流體樣本流動經過感測器及化學物質之濃度得以判定。 US 5,096,669 describes a disposable device for use with a reader device. The sample may be initially aspirated into the disposable device via capillary action and further movement of the sample within the device may be accomplished by the reader automatically pressing the balloon containing the flexible membrane within the device to cause a fluid sample The concentration of the flow through the sensor and the chemical substance is determined.

WO2013/096801描述側流偵測系統，其可包括流體特徵。在一實施方式中，描述一種盒，其包括包含層析介質及固定抗體之側流裝置，但是進一步包括藉由讀數器內之泵來致動的氣囊。對氣囊進行按壓用來使流體樣本在盒之流體通道內移動並且進入捕獲區中。在適當捕獲之後，流體樣本藉由對於氣囊之作用而被進一步推動至清洗腔室中。清洗流體之使用被描述為可沖去樣本流體之組分，諸如紅血球，該等組分可干擾偵測。然而，將側流特徵與微流控特徵組合提供一定程度的複雜性並且側流測試總體上用於定性的或半定量的量測，而非定量的量測。 WO 2013/096801 describes a lateral flow detection system that can include fluid features. In one embodiment, a cassette is described that includes a lateral flow device comprising a chromatography medium and a fixed antibody, but further comprising an air bag actuated by a pump within the reader. Pressing the balloon is used to move the fluid sample within the fluid channel of the cartridge and into the capture zone. After proper capture, the fluid sample is further pushed into the cleaning chamber by the action of the balloon. The use of a cleaning fluid is described as being a component of a sample fluid that can be flushed away, such as red blood cells, which can interfere with detection. However, combining lateral flow features with microfluidic features provides a degree of complexity and lateral flow testing is generally used for qualitative or semi-quantitative measurements, rather than quantitative measurements.

本申請人之先前申請案EP2613881描述微流控盒及相關聯讀數器。盒內之流體埠被設計來與讀數器形成不漏流體的密封以使得可藉由讀數器在盒中並且遍及整個盒來傳送氣體。 The prior application EP 2 231 881 of the Applicant describes a microfluidic cartridge and associated reader. The fluid cartridge within the cartridge is designed to form a fluid-tight seal with the reader so that the gas can be delivered by the reader in the cartridge and throughout the cartridge.

WO03/049860描述用於化學或生物化學分析之複雜裝置，其包含第一及第二層，該等第一及第二層藉由易碎的第三層分隔。在將易碎的第三層破壞之後，存在於第二層中之流體能夠進入第一層之流體網路。提供許多腔室，該等腔室必須相繼地加以壓縮以便提供各種試劑並且進行任何特定檢定。 WO 03/049860 describes a complex device for chemical or biochemical analysis comprising a first layer and a second layer separated by a fragile third layer. After destroying the fragile third layer, the fluid present in the second layer can enter the fluid network of the first layer. A number of chambers are provided which must be successively compressed to provide various reagents and to perform any particular assay.

US2015/0004680描述用於偵測樣本中之一或多個組分的生 物感測器盒。每個盒包含微流控通道、氣泵腔室及試劑泵腔室，在其上部表面包含孔隙。當相關聯感測器使用光學偵測時，亦在盒中提供清洗緩衝液。腔室被描述為單獨囊，該等囊被***該盒之凹槽中。試劑及緩衝液囊用合適液體來填充。 US 2015/0004680 describes a biosensor cartridge for detecting one or more components in a sample. Each cartridge contains a microfluidic channel, a gas pump chamber, and a reagent pump chamber containing pores on its upper surface. When the associated sensor uses optical detection, a cleaning buffer is also provided in the cartridge. The chambers are described as individual bladders that are inserted into the grooves of the box. The reagent and buffer sac are filled with a suitable liquid.

亦需要能夠更快速地並且以更小複雜性測試來自受試者之流體，諸如血液樣本。甚至需要受試者能夠自己在家進行測試。 There is also a need to be able to test fluids from a subject, such as blood samples, more quickly and with less complexity. It is even necessary for the subject to be able to test at home.

通常當受試者出現在當地醫療診所或甚至醫院時，獲取相對較大流體或血液樣本供分析並且視所執行之測試而定，可需要多個單獨小瓶或樣本。此外，在樣本收集時不進行測試的情況下，通常需要以最小化將要偵測之特定分析物的降解或損失的方式來存儲樣本。一些測試係時間敏感的並且進行測試所耗費之時間可導致疾病不當地進展，此時原本可對受試者治療。 Typically, when a subject is present at a local medical clinic or even a hospital, a relatively large fluid or blood sample is obtained for analysis and depending on the tests performed, multiple separate vials or samples may be required. In addition, where the sample is not tested at the time of sample collection, it is often desirable to store the sample in a manner that minimizes degradation or loss of the particular analyte to be detected. Some tests are time sensitive and the time it takes to perform the test can lead to an undesired progression of the disease, which can otherwise be treated to the subject.

亦存在需要受試者能夠定期地自我測試並且能夠基於測試結果來自己用藥治療的疾病及病狀。以此方面，可基於測試結果，可能使用健康照護人員的關於應服用藥物的意見對受試者進行指導。 There are also diseases and conditions that require the subject to be able to self-test on a regular basis and to be able to self-medicate based on the test results. In this regard, based on the test results, the subject may be instructed using the advice of the health care provider regarding the medication to be taken.

此外，傳統上使用臨床分析技術進行樣本之臨床分析，此等技術需要使用具有大型機器的專業實驗室進行分析。在過去幾年中，一直努力開發能夠進行此等測試的臺式大小甚至手持裝置。然而，此等設備能夠僅處理較小樣本量且/或執行各種不同類型之分析的能力係有限的。此外，若單一讀數器能夠執行各種不同組的測試，以致於為了能夠進行不同類型的測試，使用者不必具有多個不同讀數器，則係合乎需要的。 In addition, clinical analysis techniques have traditionally been used for clinical analysis of samples that require the use of specialized laboratories with large machines for analysis. In the past few years, efforts have been made to develop benchtop size and even handheld devices that can perform such tests. However, the ability of such devices to be able to process only small sample sizes and/or perform various types of analysis is limited. In addition, it is desirable if a single reader is capable of performing a variety of different sets of tests so that the user does not have to have multiple different readers in order to be able to perform different types of tests.

舉例而言，US8435738描述模組化系統，其能夠進行單一血 液樣本之多個檢定。然而，顯然提供個別及分離模組來執行不同功能並且經由樣本處理系統將樣本傳送至每個模組。亦似乎該系統包含被設計來處理樣本製備以及進行各種檢定的殼體，但是不清楚每個樣本在分析之後的後續過程為何。 For example, US8435738 describes a modular system that is capable of performing multiple assays of a single blood sample. However, it is apparent that separate and separate modules are provided to perform different functions and to transfer samples to each module via a sample processing system. It also appears that the system contains housings designed to handle sample preparation and to perform various assays, but it is not clear what the subsequent process of each sample is after analysis.

本發明部分地基於本發明人對於以下問題的研究：控制微流控盒內之液體樣本之運動以及如何在無需從盒外部引入流體的情況下，將樣本及/或清洗液結合組分從分析物偵測區盒中有效地移除，或如何使用存在於盒或相關聯讀數器中之清洗液或其他液體。發明人開發「乾式」微流控盒，其包括氣體填充腔室，該腔室可用於促進盒內之液體樣本之非常精確受控運動並且將樣本及未結合材料從盒內之分析物偵測區中移除，以使得可在氣體環境中偵測任何可偵測成份，該等成份可包括包含分析物或分析物反應產物之複合物。重要的是，本發明之盒不需要除了樣本本身以外的額外液體存在於盒中及/或引入盒中。進一步提供與盒一起使用的相關聯讀數器裝置。 The present invention is based, in part, on the inventors' study of controlling the movement of a liquid sample within a microfluidic cartridge and how to analyze the sample and/or wash fluid binding components without introducing fluid from the outside of the cartridge. The cartridge is effectively removed from the cartridge, or how the cleaning solution or other liquid present in the cartridge or associated reader is used. The inventors developed a "dry" microfluidic cartridge that includes a gas-filled chamber that can be used to facilitate very precise controlled movement of liquid samples within the cartridge and to detect samples and unbound material from analytes in the cartridge The zone is removed such that any detectable component can be detected in a gaseous environment, and such components can include a complex comprising an analyte or an analyte reaction product. Importantly, the cartridge of the present invention does not require additional liquid other than the sample itself to be present in the cartridge and/or into the cartridge. An associated reader device for use with the cartridge is further provided.

在一個方面，本發明教示方法及盒，其僅使用氣體，諸如存在於微流控盒中之腔室內之空氣，來控制盒內之液體樣本之運動以及視需要在執行偵測之前，將液體樣本及未結合材料從偵測區域中移除。 In one aspect, the present teachings teach a method and cartridge that uses only a gas, such as air present in a chamber in a microfluidic cartridge, to control the movement of a liquid sample within the cartridge and, if desired, to perform a liquid prior to performing the detection Samples and unbound material are removed from the detection area.

在另一個替代及/或補充方面，本發明提供盒、讀數器及方法，其使用單一盒及相關聯讀數器來執行多個不同檢定。 In another alternative and/or additional aspect, the present invention provides a cartridge, reader, and method that uses a single cartridge and associated reader to perform a plurality of different assays.

在第一態樣，提供用於對於液體樣本進行檢定的自給式微流控系統，該微流控系統包含： 用於接收液體樣本之樣本輸入埠，該樣本輸入埠連接至至少一個微流控通道，其中每個/該微流控通道包含置放在其中的用於進行檢定的一或多個試劑及偵測區，該偵測區用於偵測可存在於樣本中之任何分析物或其分析物反應產物；及每個/該微流控通道進一步流體連接至在每個/該偵測區下游的可壓縮、氣體填充腔室，及其中系統由三個層形成，該等層層疊在一起以界定每個/該微流控通道及該氣體填充腔室，並且其中使該腔室壓縮或減壓導致將氣體從腔室中排出或抽吸至腔室中，進而導致該/每個微流控通道內之液體樣本之運動。 In a first aspect, a self-contained microfluidic system for assaying a liquid sample is provided, the microfluidic system comprising: a sample input port for receiving a liquid sample, the sample input port being coupled to at least one microfluidic channel Each of the microfluidic channels includes one or more reagents and detection zones disposed therein for performing assays, the detection zones for detecting any analytes that may be present in the sample or An analyte reaction product; and each/the microfluidic channel is further fluidly coupled to a compressible, gas-filled chamber downstream of each/the detection zone, and wherein the system is formed of three layers that are stacked Forming each/the microfluidic channel and the gas-filled chamber together, and wherein compressing or decompressing the chamber results in expelling or pumping gas from the chamber into the chamber, thereby causing the/each Movement of a liquid sample within a microfluidic channel.

通常，但是並非排他地，系統可呈盒形式，其被設計來***相關聯讀數器裝置中。為了簡便起見，在下文涉及呈盒形式之系統，但是此不應理解為具有限制性。 Typically, but not exclusively, the system can be in the form of a box that is designed to be inserted into an associated reader device. For the sake of brevity, the following relates to a system in the form of a box, but this should not be construed as limiting.

為免生疑問，本發明不需要使用存在於盒內或與盒一起提供的液體填充囊及/或將流體(液體或氣體)從相關聯讀數器傳遞至盒的能力。在此方面，本發明之盒被視為自給式。本發明之盒在施加樣本之前係實質上無液體的並且可被視為乾燥的。在施加液體樣本之前可存在或存在於盒中之唯一流體係氣體，通常空氣。有利地，在本發明之檢定中所需要的唯一液體係液體樣本本身。 For the avoidance of doubt, the present invention does not require the ability to fill the bladder with the liquid present in or with the cartridge and/or to transfer fluid (liquid or gas) from the associated reader to the cartridge. In this regard, the cartridge of the present invention is considered to be self-contained. The cartridge of the present invention is substantially liquid free prior to application of the sample and can be considered dry. The only flow system gas, typically air, that may be present or present in the cartridge prior to application of the liquid sample. Advantageously, the only liquid system liquid sample required in the assay of the invention is itself.

在某些實施方式中，該一或多個試劑可置放在每個/該微流控通道內之第一位置中。在其他實施方式中，該一或多個試劑可置放在每個/該微流控通道內之一個以上位置中。該一或多個試劑中之至少一者可置 放在偵測區內，或替代地沒有試劑置放在偵測區內。試劑可最初以允許藉由蒸發或其他手段來乾燥的液體形式來提供。根據本發明，當該試劑最初以隨後乾燥之液體形式來提供時，術語「乾燥」應理解為意味著少於10%、5%或1%之最初液體在乾燥之後保留。 In certain embodiments, the one or more reagents can be placed in each of the first locations within the microfluidic channel. In other embodiments, the one or more reagents can be placed in more than one location within each/the microfluidic channel. At least one of the one or more reagents can be placed in the detection zone, or alternatively no reagent is placed in the detection zone. The reagents may initially be provided in a liquid form that allows drying by evaporation or other means. According to the invention, the term "drying" is understood to mean that less than 10%, 5% or 1% of the initial liquid remains after drying when the agent is initially provided in the form of a subsequently dried liquid.

在某些實施方式中，偵測區可在該一或多個試劑所置放之位置的下游。 In some embodiments, the detection zone can be downstream of where the one or more reagents are placed.

在本發明之上下文中，術語「下游」係參照將樣本施加至系統之位置及樣本流動方向。 In the context of the present invention, the term "downstream" refers to the position at which the sample is applied to the system and the direction of flow of the sample.

視需要，在液體樣本與置放在該/每個微流控通道內之該一或多個試劑反應並且將樣本及其他試劑傳遞至偵測區之後，從腔室排出之氣體視需要用來將液體從微流控通道內之偵測區移除，以使得偵測區內之任何所捕獲分析物或分析物反應產物可在實質上無液體環境中偵測。因此，在一個實施方式中，本發明提供盒及方法，其中偵測在實質上無液體環境中發生。此外，發明人觀察到僅需要使用來自腔室之對應體積之氣體將液體從偵測區中移位。因此，不需要使用可利用大量一或多種流體的習知清洗步驟，該或該等流體被設計來防止及/或最小化信號干擾。因此，有利地本發明使用低得多體積的氣體來將液體及液體內之材料從分析物偵測區中移除。此與在執行習知清洗步驟方面所理解的概念係顯著不同的。 If desired, after the liquid sample reacts with the one or more reagents placed in the/each microfluidic channel and the sample and other reagents are delivered to the detection zone, the gas exhausted from the chamber is used as needed The liquid is removed from the detection zone within the microfluidic channel such that any captured analyte or analyte reaction product within the detection zone can be detected in a substantially liquid free environment. Accordingly, in one embodiment, the present invention provides a cartridge and method wherein detection occurs in a substantially liquid-free environment. Furthermore, the inventors observed that it is only necessary to use a gas from a corresponding volume of the chamber to displace the liquid from the detection zone. Thus, there is no need to use conventional cleaning steps that can utilize a large number of one or more fluids that are designed to prevent and/or minimize signal interference. Thus, advantageously, the present invention uses a much lower volume of gas to remove material from the liquid and liquid from the analyte detection zone. This is significantly different from the concept system understood in performing the conventional cleaning steps.

微流控盒可包含複數個微流控通道，其中複數個微流控通道全部與單一樣本輸入埠流體連通。根據本發明，樣本埠可與劃分成該等複數個微流控通道之微流控通道連通。該等複數個微流控通道中之每一者可與相應氣體填充腔室流體連通，或兩個或兩個以上微流控通道可與單一氣 體填充腔室流體連通。根據本發明，每個腔室可個別地或獨立地控制，從而允許獨立控制個別微流控通道內之樣本之運動，或可同時控制複數個該微流控通道中之樣本之運動。在一些實施方式中，除了與盒中之一或多個微流控通道流體連通以外，該氣體填充腔室不與可存在於微流控盒或相關聯讀數器中之任何其他特徵連接。舉例而言，僅來自氣體填充腔室之開口/出口可為與該微流控通道之開口。因此，就不具有閥、埠或以其他方式與盒外部連通而言，該氣體填充腔室可為密封的。在一實施方式中，樣本輸入埠連接至每個/該微流控通道之第一端並且每個/該微流控通道之第二端連接至該等氣體填充腔室中之一或多者之該開口。在此實施方式中，氣體填充腔室可被視為在樣本輸入埠之下游並且在每個/該微流控通道的相對於樣本輸入埠之相反端。 The microfluidic cartridge can include a plurality of microfluidic channels, wherein the plurality of microfluidic channels are all in fluid communication with a single sample input port. According to the present invention, the sample file can be in communication with a microfluidic channel divided into the plurality of microfluidic channels. Each of the plurality of microfluidic channels can be in fluid communication with a respective gas filled chamber, or two or more microfluidic channels can be in fluid communication with a single gas filled chamber. In accordance with the present invention, each chamber can be individually or independently controlled to allow for independent control of the motion of samples within an individual microfluidic channel, or to simultaneously control the motion of a plurality of samples in the microfluidic channel. In some embodiments, the gas-filled chamber is not coupled to any other feature that may be present in the microfluidic cartridge or associated reader, except that it is in fluid communication with one or more of the microfluidic channels in the cartridge. For example, only the opening/outlet from the gas filled chamber can be the opening to the microfluidic channel. Thus, the gas-filled chamber can be sealed without having a valve, a weir, or otherwise communicating with the exterior of the cartridge. In one embodiment, a sample input port is coupled to each of the first ends of the microfluidic channel and a second end of each/the microfluidic channel is coupled to one or more of the gas filled chambers The opening. In this embodiment, the gas-filled chamber can be considered to be downstream of the sample input port and at the opposite end of each/the microfluidic channel relative to the sample input port.

除非上下文另外規定，否則術語「流體連通」應理解為意味著包括氣體或液體之流體能夠在相關部分之間傳送。 Unless the context dictates otherwise, the term "fluid communication" is understood to mean that a fluid comprising a gas or liquid can be transferred between relevant parts.

視需要，微流控盒可進一步包含一或多個吸收器特徵，其被設計來接收流體廢物及/或過量液體樣本。為免生疑問，本發明之一些實施方式特定地排除一或多個吸收器特徵，其可為有利的。 If desired, the microfluidic cartridge can further include one or more absorber features designed to receive fluid waste and/or excess liquid sample. For the avoidance of doubt, some embodiments of the invention specifically exclude one or more absorber features, which may be advantageous.

本發明之盒設計可容易地調適以便執行許多不同檢定並且由此可被視為各種類似及/或不同檢定之檢定平臺。盒及安置在其中之通道可以審閱本說明書之熟習此項技術者已知的任何方式來形成，其可包括微影術、濕化學蝕刻、雷射燒蝕、注射模造、模頭衝壓、壓花及印刷技術。根據本發明之第一態樣，盒及安置在其中之通道及其他特徵藉由三個分離基板，亦即第一、第二及第三基板之層疊物來形成，諸如頂部及底部基板 與安置在頂部與底部基板之間的中間基板。三個層可藉由施加熱量或使用黏著劑來密封在一起。此外，中間層可本身呈黏著劑層形式，其能夠黏附頂部及底部基板。 The cartridge design of the present invention can be easily adapted to perform a number of different assays and thus can be viewed as a variety of similar and/or different assays. The cartridge and the passageway disposed therein can be formed by any means known to those skilled in the art, including lithography, wet chemical etching, laser ablation, injection molding, die stamping, embossing. And printing technology. According to a first aspect of the invention, the cassette and the channels and other features disposed therein are formed by three separate substrates, namely a stack of first, second and third substrates, such as top and bottom substrates and placement An intermediate substrate between the top and bottom substrates. The three layers can be sealed together by applying heat or using an adhesive. In addition, the intermediate layer may itself be in the form of an adhesive layer that is capable of adhering the top and bottom substrates.

在一實施方式中，三個基板係平面的。通常，第一及第三(例如，頂部及底部)及視需要第二(例如，中間)基板在性質上係實質上均勻的。亦即，該基板之厚度係均勻的並且在基板之表面上係不變化的。 In one embodiment, the three substrates are planar. Typically, the first and third (eg, top and bottom) and, if desired, the second (eg, intermediate) substrate are substantially uniform in nature. That is, the thickness of the substrate is uniform and does not change on the surface of the substrate.

在具體實施方式中，底部基板黏附至中間基板，該中間基板具有已經安置在其中之通道。在底部基板上，將執行檢定所需要之試劑置放在其特定置放區中，並且一旦藉由樣本來重構，該等試劑藉由通道壁(藉由中間黏著劑層來形成)來保持在原位並且藉由印刷在底部基板上之特徵，例如疏水性油墨來防止在整個所形成通道中擴散。以此方式，藉由所有四個側面上之特徵來防止試劑擴散至其置放區外部。然後，試劑乾燥及最終頂層基板黏附中間層產生完全形成盒。將檢定試劑置放盒中之許多其他方法可藉由熟習此項技術者來設想。 In a specific embodiment, the bottom substrate is adhered to an intermediate substrate having a channel that has been disposed therein. On the bottom substrate, the reagents required to perform the assay are placed in their particular placement area, and once reconstituted by the sample, the reagents are retained by the channel walls (formed by the intermediate adhesive layer). Dispersion throughout the formed channel is prevented in situ and by features printed on the bottom substrate, such as hydrophobic ink. In this way, the diffusion of the reagent to the outside of its placement zone is prevented by the features on all four sides. The reagent is then dried and the final top substrate adheres to the intermediate layer to produce a fully formed cell. Many other methods of placing assay reagents in a cartridge can be envisioned by those skilled in the art.

本發明之盒可藉由在此項技術中已知之捲筒或卷對卷過程從可撓性聚合物膜、塑膠或金屬箔之輥來形成。 The cartridge of the present invention can be formed from a roll of flexible polymeric film, plastic or metal foil by a roll or roll-to-roll process known in the art.

有利地，發明人發現當盒由三個單獨平面基板之層疊物形成，盒之頂部及底部基板不必具有不同厚度及/或具有不同厚度之部分或由其他材料形成。因此，頂部及底部層可具有均勻厚度並且由單一材料形成。此簡化盒之製造及相關聯成本。用於以形成頂部及視需要底部層之材料可為可撓性的，但是在通道及氣體腔室尺度下，其為相當堅硬的，但是稍微具有彈性。意外地，形成該/每個氣體填充腔室之頂部及底部表面，尤其頂 部外表面的基板可為彈性的，即使在基板之整個表面上，頂部表面之厚度係均勻的亦如此。 Advantageously, the inventors have discovered that when the cartridge is formed from a laminate of three separate planar substrates, the top and bottom substrates of the cartridge need not have portions of different thicknesses and/or have different thicknesses or be formed from other materials. Thus, the top and bottom layers can have a uniform thickness and be formed from a single material. The manufacture of this simplified box and associated costs. The material used to form the top and optionally the bottom layer may be flexible, but at the channel and gas chamber dimensions, it is relatively stiff, but slightly elastic. Surprisingly, the substrate forming the top and bottom surfaces of the/each gas-filled chamber, particularly the top outer surface, can be resilient, even if the thickness of the top surface is uniform over the entire surface of the substrate.

可用於將層密封在一起的黏著劑亦可組合以促進每個/該腔室之可壓縮性。因此，腔室之可壓縮性質可部分地歸因於黏著劑係彈性的，以及頂部及視需要底部基板係彈性的。與先前技術相反，本發明之氣體填充腔室不需要包含形成腔室之外表面的可撓性膜或片材，與用於形成頂部及視需要底層之其餘部分的基板相比，該腔室之外表面由不同材料製成及/或具有不同厚度或可撓性。因此，頂層及視需要底層由在整個層上具有均勻厚度之單一材料製成。理想地，頂部及底部層由相同材料製成並且具有均勻厚度。此簡化盒之製造，考慮到成本方面，此係至關重要的。 Adhesives that can be used to seal the layers together can also be combined to promote the compressibility of each/the chamber. Thus, the compressible nature of the chamber can be attributed in part to the elasticity of the adhesive, as well as the top and, if desired, the bottom substrate being elastic. In contrast to the prior art, the gas-filled chamber of the present invention need not include a flexible film or sheet that forms the outer surface of the chamber, as compared to the substrate used to form the top and optionally the remainder of the bottom layer. The outer surface is made of a different material and/or has a different thickness or flexibility. Thus, the top layer and optionally the bottom layer are made of a single material having a uniform thickness throughout the layer. Ideally, the top and bottom layers are made of the same material and have a uniform thickness. The manufacture of this simplified box is critical in terms of cost.

盒可由任何合適材料形成，諸如聚碳酸酯、聚烯烴，諸如環狀石蠟共聚物(COCs)、聚酯、聚苯乙烯、PMMA等，並且該/每個基板可由單一或複數個材料形成。在包含三個基板之實施方式中，中間基板包含貫穿基板切割之圖案，其對應於盒之某些特徵，諸如該通道、氣體填充腔室、廢物吸收器等。藉由施加並層疊(諸如藉由熱封、膠合、吻合等)經適當切割之頂部及底部基板，以將中間基板層疊在頂部與底部基板之間，可提供其中安置通道及其他特徵的盒。每個層可個別地提供並且夾在一起，或三個層可彼此連接並且藉由將該等層彼此疊置折疊來形成層疊物以便形成盒。頂部及/或底部基板可從不同於中間基板的材料形成或用該材料來塗布且/或將黏著劑材料施加至基板中之任一者以促進三個基板黏在一起。頂部及/或底部基板中之特徵可被設計來與讀數器裝置中之特徵共置(如在下文論述)，從而可促進讀數器中之盒之正確定位。 The cartridge may be formed from any suitable material, such as polycarbonate, polyolefin, such as cyclic paraffin copolymers (COCs), polyester, polystyrene, PMMA, etc., and the/each substrate may be formed from a single or multiple materials. In an embodiment comprising three substrates, the intermediate substrate comprises a pattern cut through the substrate that corresponds to certain features of the cartridge, such as the channel, gas filled chamber, waste absorber, and the like. A cassette in which the channels and other features are disposed can be provided by applying and laminating (such as by heat sealing, gluing, stapling, etc.) the appropriately cut top and bottom substrates to laminate the intermediate substrate between the top and bottom substrates. Each layer may be provided separately and sandwiched together, or the three layers may be connected to each other and formed by stacking the layers on top of one another to form a laminate. The top and/or bottom substrate may be formed from or coated with a material other than the intermediate substrate and/or any one of the adhesive materials applied to the substrate to facilitate bonding of the three substrates together. Features in the top and/or bottom substrate can be designed to co-locate with features in the reader device (as discussed below) to facilitate proper positioning of the cartridge in the reader.

在一便利實施方式中，本發明之檢定之讀出被設計成以光學方式來偵測。在此方面，相關聯讀數器包括光學偵測構件，諸如光譜儀或螢光計，其被設計來偵測從盒之偵測區域發出之電磁輻射。對於螢光偵測，讀數器內之光譜儀或螢光計偵測從偵測區內之材料發出之螢光。因此，被設計成面對讀數器中之光譜儀或螢光計的盒之第一或第三層(例如，頂部或底部層)之至少一部分必須在電磁輻射譜之合適範圍中係光學可透射的。在螢光偵測的情況下，盒之第一或第三(例如，頂部或底部)層之至少一部分必須在涵蓋激發波長及偵測波長之範圍中係光學可透射的。舉例而言，盒之第一及第三(例如，頂部或底部)層之至少一部分必須在200-1200nm範圍中係光學可透射的。 In a convenient embodiment, the readout of the assay of the present invention is designed to be optically detected. In this regard, the associated reader includes an optical detection component, such as a spectrometer or fluorometer, designed to detect electromagnetic radiation emanating from the detection area of the cartridge. For fluorescence detection, the spectrometer or fluorometer in the reader detects the fluorescence emitted from the material in the detection zone. Thus, at least a portion of the first or third layer (eg, the top or bottom layer) of the cartridge designed to face the spectrometer or fluorometer in the reader must be optically transmissive in the proper range of electromagnetic radiation spectrum. . In the case of fluorescent detection, at least a portion of the first or third (eg, top or bottom) layer of the cartridge must be optically transmissive in the range encompassing the excitation wavelength and the detection wavelength. For example, at least a portion of the first and third (eg, top or bottom) layers of the cartridge must be optically transmissive in the range of 200-1200 nm.

當第一及第三(例如，頂部及/或底部)層由單一材料製成時，應認識到整個第一及第三(例如，頂部及/或底部)層係光學可透射的並且並非僅其一部分。然而，油墨及/或遮罩可用於防止或最小化合適波長之電磁輻射濾出或散射至偵測區外部。在一實施方式中，涵蓋偵測區或其一部分的第三(例如，底部)層之一部分可用材料塗布，該材料被設計成最大化任何螢光信號朝向讀數器之光學偵測構件的發射。 When the first and third (eg, top and/or bottom) layers are made of a single material, it should be recognized that the entire first and third (eg, top and/or bottom) layers are optically transmissive and not only Part of it. However, the ink and/or mask can be used to prevent or minimize the filtering or scattering of electromagnetic radiation of a suitable wavelength outside of the detection zone. In one embodiment, a portion of the third (eg, bottom) layer that encompasses the detection zone or a portion thereof may be coated with a material that is designed to maximize the emission of any fluorescent signal toward the optical detection component of the reader.

第一及第三(例如，頂部及底部)基板可藉由鉸鏈來連接，該鉸鏈允許兩個基板彼此相鄰折疊，並且中間基板安置在其之間。或者鉸鏈可提供在第一與第二(例如，頂部與中間)基板以及第二與第三(例如，中間與底部)基板之間，以使得第一、第二及第三(例如，頂部、中間及底部)基板彼此相鄰折疊並且可從基板之單一片材形成。 The first and third (eg, top and bottom) substrates may be joined by a hinge that allows the two substrates to be folded adjacent to each other with the intermediate substrate disposed therebetween. Or a hinge may be provided between the first and second (eg, top and middle) substrates and the second and third (eg, middle and bottom) substrates such that the first, second, and third (eg, top, The middle and bottom substrates are folded adjacent to each other and can be formed from a single sheet of the substrate.

重要的是，該氣體填充腔室被設計來與讀數器中之一或多個 特徵並置，該等特徵被設計來接觸該氣體填充腔室之外表面(亦即，呈三個基板層疊物形式之頂部及/或底部基板，其以實質上水平方式提供至讀數器)並且能夠控制施加至腔室之外表面/從腔室之外表面移除之力，例如，壓縮。向該/每個腔室之外表面施加力導致腔室變形並且腔室內之氣體從腔室排出至微流控通道中。相反地，隨後將施加至該/每個腔室之力減少例如，減壓，導致腔室更小變形並且視需要返回到未變形狀態，以使得氣體從微流控通道抽吸回到腔室中。 Importantly, the gas-filled chamber is designed to be juxtaposed with one or more features in the reader, the features being designed to contact the outer surface of the gas-filled chamber (ie, in the form of three substrate laminates) The top and/or bottom substrate is provided to the reader in a substantially horizontal manner and is capable of controlling the force applied to/from the outer surface of the chamber, for example, compression. Applying a force to the outer surface of the/each chamber causes the chamber to deform and the gas within the chamber is discharged from the chamber into the microfluidic channel. Conversely, the force applied to the/each chamber is then reduced, for example, to a reduced pressure, resulting in a smaller deformation of the chamber and returning to an undeformed state as needed to allow gas to be drawn back from the microfluidic channel back into the chamber. in.

應認識到在不施加力的情況下，氣體填充腔室通常包含最大體積之氣體。在施加力後，氣體從氣體填充腔室中排出，由此減少腔室內之氣體之體積。隨後減少施加至腔室之力允許氣體抽吸回到腔室中，從而導致氣體填充腔室內之氣體之體積增加。 It will be appreciated that the gas filled chamber typically contains a maximum volume of gas without application of force. After the force is applied, the gas is expelled from the gas-filled chamber, thereby reducing the volume of gas within the chamber. Subsequent reduction of the force applied to the chamber allows gas to be drawn back into the chamber, resulting in an increase in the volume of gas within the gas-filled chamber.

每個腔室內之氣體通常為空氣，但是可引入其他氣體或氣體混合物。舉例而言，若置放在每個/該微流控通道內之試劑中之任一者可能氧化或另外在存在於空氣中時具有較短壽命，盒及相關聯通道及腔室可用惰性氣體諸如氮等來填充。總體上涉及作為空氣之氣體，但是此不應被視為具有限制性。 The gas in each chamber is typically air, but other gases or gas mixtures can be introduced. For example, if any of the reagents placed in each/the microfluidic channel may oxidize or otherwise have a shorter lifetime when present in air, the cartridge and associated channels and chambers may be inert. Filled with such as nitrogen. It is generally referred to as a gas for air, but this should not be considered limiting.

通常在使用中，在樣本施加之前，可將盒提供至或***讀數器中並且將力施加至該/每個腔室以使氣體從該/每個腔室中排出並且進入該/每個微流控通道中。盒可被視為「準備好」用於樣本施加。 Typically in use, prior to application of the sample, the cartridge can be provided to or inserted into the reader and a force applied to the/each chamber to allow gas to escape from the/each chamber and into the/each micro In the flow control channel. The box can be considered "ready" for sample application.

樣本，諸如樣本血液，或任何其他液體樣本可經由樣本輸入埠來引入盒中。樣本可藉由使樣本與輸入埠接觸來直接引入。或者，樣本可首先使用樣本收集構件來收集並且此等樣本收集構件，諸如浸量尺、微 量吸移管、毛細管等與樣本輸入埠接觸或***樣本輸入埠中以使得可將樣本引入盒及微流控通道中。在一些實施方式中，諸如在執行核酸分析時，可需要在封閉系統中執行任何分析。因此，被設計來將樣本引入微流控盒中之樣本收集構件可滿足雙重目的：引入樣本及一旦樣本收集構件與樣本輸入埠接觸或***樣本輸入埠中，即將樣本輸入埠密封。 A sample, such as sample blood, or any other liquid sample, can be introduced into the cassette via a sample input port. The sample can be introduced directly by contacting the sample with the input port. Alternatively, the sample may first be collected using a sample collection member and such sample collection members, such as dipsticks, micropipettes, capillaries, etc., are contacted with the sample input port or inserted into the sample input port such that the sample can be introduced into the cassette and the microfluidic In the control channel. In some embodiments, such as when performing nucleic acid analysis, any analysis may need to be performed in a closed system. Thus, a sample collection member designed to introduce a sample into a microfluidic cartridge can serve a dual purpose: to introduce a sample and to seal the sample input 一旦 once the sample collection member is in contact with or inserted into the sample input port.

在樣本與盒之樣本輸入埠接觸或引入該樣本輸入埠中之後，樣本可最初經由毛細管作用來抽吸至微流控通道中。或者可將樣本主動地抽吸至微流控通道中，方法係藉由減少施加至該/每個腔室之力以使得氣體被抽吸至腔室中，進而將液體樣本至該/每個微流控通道中並且沿著微流控通道抽吸。 After the sample is brought into contact with or introduced into the sample input port, the sample can be initially drawn into the microfluidic channel via capillary action. Alternatively, the sample can be actively pumped into the microfluidic channel by reducing the force applied to the/each chamber such that gas is drawn into the chamber, thereby passing the liquid sample to the/each Suction in the microfluidic channel and along the microfluidic channel.

在一實施方式中，液體樣本最初沿著單一微流控通道抽吸，該微流控通道劃分成複數個微流控通道，該等複數個通道中之每一者能夠執行一或多個檢定。以此方式，可提供單一樣本，該樣本進而劃分成多個部分或等分試樣。 In one embodiment, the liquid sample is initially drawn along a single microfluidic channel, the microfluidic channel being divided into a plurality of microfluidic channels, each of the plurality of channels being capable of performing one or more assays . In this way, a single sample can be provided which is in turn divided into a plurality of sections or aliquots.

一旦諸如經由毛細管作用及/或主動地抽吸樣本穿過盒，將樣本引入盒及該/每個微流控通道中，謹慎地控制/促進在盒及相關聯通道內以及在整個盒及相關聯通道中之進一步流體傳送，方法係經由控制施加至該/每個氣體腔室之壓力，導致將氣體引入該/每個氣體填充腔室中及/或從氣體填充腔室中排出。抽吸回到該/每個腔室中之氣體通常由於真空效應用來將液體樣本沿著該/每個微流控通道朝向該/每個腔室抽吸，並且從該/每個腔室排出之氣體將該/每個微流控通道內之液體推動遠離該/每個腔室，到達輸入埠並且視需要進入可能存在之流體廢物吸收器中。 Once the sample is introduced into the cartridge and the/each microfluidic channel, such as via capillary action and/or actively pumping the sample through the cartridge, carefully control/promote within the cartridge and associated channels and throughout the cartridge and related Further fluid transfer in the associated passage is accomplished by controlling the pressure applied to the/each gas chamber, causing gas to be introduced into and/or from the gas filled chamber. The gas drawn back into the/each chamber is typically used to draw a liquid sample along the/each microfluidic channel toward the/each chamber due to a vacuum effect, and from the/each chamber The vented gas pushes the liquid within each microfluidic channel away from the/each chamber, reaches the input port and, as needed, enters a fluid waste absorber that may be present.

如上所述，吸收器特徵係完全任擇的。根據本發明，可經由適當流體控制及氣體腔室管理來確保一旦樣本引入盒中，樣本或其他液體不能從樣本埠中排出。在樣本施加之前，每個/該腔室可壓縮至最大程度以使得不可將樣本從樣本輸入埠排出。有利地，在樣本施加之前壓縮每個/該氣體填充腔室意味著在進行任何檢定並釋放每個/該氣體填充腔室上之任何壓縮壓力之後，液體樣本被進一步抽吸至盒中並且可能進入氣體填充腔室中，遠離樣本輸入埠。從在檢定之後，將任何樣本與使用者分離的角度來講，此可被視為有用的安全特徵。 As mentioned above, the absorber characteristics are completely optional. In accordance with the present invention, it is ensured through proper fluid control and gas chamber management that once the sample is introduced into the cartridge, the sample or other liquid cannot be discharged from the sample cartridge. Each/the chamber can be compressed to a maximum extent prior to sample application such that the sample cannot be discharged from the sample input port. Advantageously, compressing each/the gas-filled chamber prior to sample application means that after any verification is performed and any compression pressure on each/the gas-filled chamber is released, the liquid sample is further drawn into the cartridge and possibly Enter the gas-filled chamber away from the sample input port. This can be considered a useful security feature from the standpoint of separating any sample from the user after verification.

流體運動可藉由讀數器中之力控制構件來非常精確地控制。此外，每個通道內之流體之位置可視需要藉由讀數器、藉由沿著微流控通道定位之構件諸如電極來偵測，該等構件與讀數器接觸並且可回饋每個/該微流控通道中之任何液體及/或流體之位置，從而允許讀數器經由向氣體/空氣填充腔室施加力/壓力來非常謹慎地判定流體運動之位置及/或速率。 Fluid motion can be controlled very precisely by the force control members in the reader. In addition, the position of the fluid within each channel can be detected by a reader, by means of components positioned along the microfluidic channel, such as electrodes, which are in contact with the reader and can feed back each/the microflow The position of any liquid and/or fluid in the channel is controlled to allow the reader to very carefully determine the position and/or velocity of the fluid motion by applying a force/pressure to the gas/air filling chamber.

如確認，在使用中，樣本經由樣本輸入埠諸如經由受試者/患者或其他構件例如吸移管、毛細管等之直接接觸來施加至盒。在一個較佳實施方式中，樣本輸入埠係盒之側面或端面(例如，頂部端面)中之孔洞。合意地，盒可呈包含頂部及底部端面及四個邊緣之總體上薄平面裝置形式。在此佈置中，樣本輸入埠可在盒之一個邊緣中或在頂部端面上形成，以使得使用者僅需要將樣本與在邊緣中或在頂部端面上形成之孔洞接觸，以便使得樣本能夠吸收至盒中。在使用中，使用者將流體樣本與埠/孔洞接觸並且在某些實施方式中，歸因於盒內之該通道之尺寸，流體藉由毛細管作用來抽吸至盒中。樣本埠/孔洞之尺寸可小於通道之尺寸。因此，在將流 體從該/每個微流控通道排出時，任擇流體廢物吸收器提供較大空隙區域，可將廢液樣本及任何未反應的試劑/標記物朝向該空隙區域引導並且進入廢物吸收器，而不是穿過樣本輸入埠。 As confirmed, in use, the sample is applied to the cartridge via sample input, such as via direct contact with a subject/patient or other member such as a pipette, capillary, or the like. In a preferred embodiment, the sample is input into a hole in the side or end face (e.g., the top end face) of the tether box. Desirably, the cartridge can be in the form of a generally thin planar device comprising top and bottom end faces and four edges. In this arrangement, the sample input port can be formed in one edge of the cartridge or on the top end face such that the user only needs to contact the sample with a hole formed in the edge or on the top end face to enable the sample to be absorbed into In the box. In use, the user contacts the fluid sample with the helium/hole and, in some embodiments, the fluid is drawn into the cartridge by capillary action due to the size of the passage within the cartridge. The size of the sample 埠/hole can be smaller than the size of the channel. Thus, when fluid is expelled from the/each microfluidic channel, the optional fluid waste absorber provides a larger void area, and the waste sample and any unreacted reagents/markers can be directed toward the void region and enter Instead of passing through the sample input 埠, the waste absorber.

在某些實施方式中，不提供廢物吸收器。因為不需要移除樣本，或僅需要從該/每個微流控通道內之偵測區移除樣本及未反應的試劑/標記物。可實現流體運動之謹慎控制，以使得從該/每個氣體填充腔室排出之氣體足以將樣本及任何未反應的/不必要的試劑/標記物從偵測區中完全移除掉。在液體樣本施加之前，該氣體填充腔室之初始最大壓縮確保不可推動樣本超過樣本輸入埠。微流控通道內之流體運動之此謹慎控制意味著可不需要廢物吸收器及/或較大體積之清洗流體，導致簡化製造/使用及成本節約。 In certain embodiments, no waste absorber is provided. Because there is no need to remove the sample, or only the sample and unreacted reagents/markers need to be removed from the detection zone within the/each microfluidic channel. Careful control of fluid motion can be achieved such that the gas exiting the/each gas-filled chamber is sufficient to completely remove the sample and any unreacted/unnecessary reagents/markers from the detection zone. The initial maximum compression of the gas-filled chamber prior to application of the liquid sample ensures that the sample cannot be pushed beyond the sample input. This careful control of fluid movement within the microfluidic channel means that no waste absorbers and/or larger volumes of cleaning fluid may be required, resulting in simplified manufacturing/use and cost savings.

盒中之該微流控通道亦可包含一或多個流體止擋特徵，其被設計來防止樣本藉助於僅毛細管作用而經過止擋特徵。亦即，必須藉由用來抽吸或推動盒內之液體樣本之力的作用來主動地迫使樣本經過該止擋特徵及/或進一步沿著該微流控通道，諸如作用於氣體填充腔室之壓縮及/或減壓力。止擋特徵可為疏水性材料(例如，可印刷的導電或非導電油墨)或改變通道表面之表面性質，由此產生親水性/疏水性差異的過程或材料(例如，經由雷射燒蝕、表面刻痕、表面材料移除、蒸發金屬材料等)，其被設計來鄰接或作為壁特徵或塗布在通道之壁(例如，頂部、側面及/或底部)上。在通道藉助於三個基板層疊在一起從而形成通道來形成的一實施方式中，疏水性材料可施加至頂部及/或底部基板，以使得當三個基板層疊在一起時，疏水性止擋材料形成該通道之頂部及/或底部表面上之特徵(該通道之壁藉由中 間層來形成)。替代地或另外，可與通道相鄰或在通道內提供較小單向通氣孔，該通氣孔能夠允許空氣排放至盒外部或盒內之空隙，但是不允許空氣或液體進入該微流控通道。藉由毛細管作用進入盒之液體填充至通氣孔但是在不施加額外力的情況下不超過它。 The microfluidic channel in the cartridge may also include one or more fluid stop features designed to prevent the sample from passing through the stop feature by capillary action alone. That is, the sample must be actively forced to pass the stop feature and/or further along the microfluidic channel by acting as a force for pumping or pushing a liquid sample within the cartridge, such as acting on a gas filled chamber Compression and / or pressure reduction. The stop feature can be a hydrophobic material (eg, a printable conductive or non-conductive ink) or a process or material that changes the surface properties of the channel surface, thereby creating a hydrophilic/hydrophobic difference (eg, via laser ablation, Surface scoring, surface material removal, evaporating metal materials, etc.) are designed to abut or be wall features or coated on the walls of the channel (eg, top, side, and/or bottom). In one embodiment in which the channels are formed by laminating three substrates together to form channels, a hydrophobic material can be applied to the top and/or bottom substrate such that when the three substrates are stacked together, the hydrophobic stop material The features on the top and/or bottom surface of the channel are formed (the walls of the channel are formed by an intermediate layer). Alternatively or additionally, a small one-way vent may be provided adjacent to or within the channel that is capable of allowing air to be vented to the exterior of the cartridge or to the void within the cartridge, but does not allow air or liquid to enter the microfluidic passage . The liquid entering the cartridge by capillary action fills the vent but does not exceed it without applying additional force.

在提供多個通道以便執行分開及/或重複檢定的一實施方式中，流體止擋特徵可在樣本輸入埠之下游提供在每個通道中。以此方式，樣本最初經由樣本輸入埠進入盒，但是藉由流體止擋特徵來防止填充每個微流控通道之長度。為了開始每個檢定，必須藉由將氣體抽吸回到該/每個氣體腔室中來將樣本主動地抽吸經過流體止擋物及沿著每個微流控通道以便接觸該一或多個試劑。有利地，此確保每個檢定可根據需要同時或在不同時間開始並且亦用來最大限度地減少可由於樣本差異而出現的問題，諸如血液血球比容值及進而例如黏度差異。 In one embodiment in which multiple channels are provided to perform separate and/or repeated assays, fluid stop features may be provided in each channel downstream of the sample input port. In this way, the sample initially enters the cartridge via the sample input, but the length of each microfluidic channel is prevented from filling by the fluid stop feature. To begin each assay, the sample must be actively drawn through the fluid stop and along each microfluidic channel to contact the one or more by pumping gas back into the/each gas chamber. Reagents. Advantageously, this ensures that each assay can be initiated simultaneously or at different times as needed and also to minimize problems that can arise due to sample differences, such as blood hematocrit values and thus, for example, viscosity differences.

亦較佳止擋特徵定位在可能存在之流體廢物吸收器之上游，以使得在最初施加後，樣本不流至廢物腔室中。當向該/每個氣體填充腔室施加足夠力以便主動地推動該/每個通道內之液體樣本時，液體可經過流體廢物吸收器上游之止擋特徵並且進入廢物吸收器。此止擋特徵亦可以一定方式來設計，該方式使得雖然其在最初接觸時防止流體進入吸收器，但是樣本可最終濕潤越過此止擋特徵並且流至吸收器中，但是僅一旦樣本填充樣本通道時才會如此。一旦此等通道變滿，止擋特徵上之毛細管力增加並且過量樣本可流動越過止擋特徵並且進入吸收器中。以此方式，吸收器可充當過量樣本施加之溢流並且流體止擋特徵可充當定時閘門，控制此液體運動。在其他實施方式中，此止擋特徵並非必需的。吸收器可用樣本 填充並且充當儲槽，可藉由減少氣體填充腔室上之力來從該儲槽中抽吸樣本以便將此樣本從吸收器傳遞至樣本通道中。 It is also preferred that the stop feature be positioned upstream of the fluid waste absorber that may be present such that after initial application, the sample does not flow into the waste chamber. When sufficient force is applied to the/each gas filling chamber to actively propel the liquid sample within the/each channel, the liquid can pass through the stop feature upstream of the fluid waste absorber and into the waste absorber. This stop feature can also be designed in such a way that although it prevents fluid from entering the absorber upon initial contact, the sample can eventually wet over the stop feature and flow into the absorber, but only once the sample fills the sample channel This will be the case. Once these channels become full, the capillary force on the stop feature increases and the excess sample can flow past the stop feature and into the absorber. In this way, the absorber can act as an overflow for excess sample application and the fluid stop feature can act as a timing gate to control this liquid motion. In other embodiments, this stop feature is not required. The absorber can be filled with a sample and act as a reservoir from which a sample can be aspirated by reducing the force on the gas-filled chamber to transfer this sample from the absorber into the sample channel.

在一實施方式中，廢物吸收器被設計成盒之空隙區域，耗盡的流體/樣本或不需要或被視為不希望有的流體可排空至該空隙區域中。舉例而言，全血含有許多蛋白質及其他因子，該等因子可干擾檢定反應及/或經由例如螢光偵測來偵測所捕獲的分析物。本發明允許執行樣本(例如，全血)內之任何分析物的最初結合及/或反應，但是所有或實質上所有存在於液體樣本中之未結合材料及反應之後的剩餘液體可隨後從偵測區排空，視需要至廢物腔室，使得能夠在實質上無液體或氣體環境中執行進一步反應及/或偵測。 In one embodiment, the waste absorber is designed as a void region of the cartridge, and the depleted fluid/sample or fluid that is not required or deemed undesirable may be emptied into the void region. For example, whole blood contains a number of proteins and other factors that can interfere with the assay reaction and/or detect the captured analyte via, for example, fluorescence detection. The present invention allows for the initial binding and/or reaction of any analyte within a sample (eg, whole blood), but all or substantially all of the unbound material present in the liquid sample and the remaining liquid after the reaction can be subsequently detected The emptying of the zone, as needed to the waste chamber, enables further reaction and/or detection to be performed in a substantially liquid or gaseous environment.

然而，如上所述，可不需要包括廢物吸收器。有利地，本發明人觀察到在向該/每個氣體腔室施加力時從該/每個氣體腔室排出之氣體足以推動/傳送液體樣本及未結合/未反應的材料遠離偵測區。因此，僅被捕獲、結合或固定材料保持實質上無液體環境中之偵測區中，並且有利地，容易地進行任何此材料之偵測。 However, as noted above, it may not be necessary to include a waste absorber. Advantageously, the inventors have observed that the gas exiting the gas chamber is sufficient to push/deliver the liquid sample and unbound/unreacted material away from the detection zone when a force is applied to the/each gas chamber. Thus, only the captured, bonded or immobilized material remains in the detection zone in a substantially liquid free environment, and advantageously any such material is readily detected.

除了每個/該微流控通道以外，本發明之盒可包含一或多個電極特徵，其與該通道接觸並且由此與一旦引入盒中之樣本接觸。電極被設計來接觸讀數器內之電氣觸點，使得能夠在適當情況下獲取各種讀數。舉例而言，盒中之一或多個電極可被設計來偵測盒之正確負載並且讀數器可以信號告知使用者是否盒a)正確***讀數器中及/或b)樣本正確地負載至盒中，例如直至流體止擋特徵。電極亦可對於樣本本身執行一或多個電氣量測。舉例而言，在樣本係全血樣本時，電極可進行樣本之血球比容量測， 此在判定將要偵測之分析物之精確濃度中可為至關重要的。導電率及/或阻抗量測可取決於所研究之樣本來判定。因此，本發明之盒可不僅經由與任何分析物之結合/反應來偵測是否分析物存在於樣本中，而且亦可進行樣本之電氣量測。電極亦可用於確認藉由從氣體填充腔室排出之氣體來從偵測區域移除樣本已經正確地發生，因為當液體存在或不存在時偵測到之電導率會顯著變化。亦可向氣體填充腔室提供電極以便以信號告知每個/該腔室之壓縮度。 In addition to each/the microfluidic channel, the cartridge of the present invention can include one or more electrode features that are in contact with the channel and thereby contact the sample once introduced into the cartridge. The electrodes are designed to contact the electrical contacts within the reader so that various readings can be taken where appropriate. For example, one or more of the electrodes in the cartridge can be designed to detect the correct load of the cartridge and the reader can signal the user whether the cartridge a) is properly inserted into the reader and/or b) the sample is properly loaded into the cartridge Medium, for example up to the fluid stop feature. The electrodes may also perform one or more electrical measurements on the sample itself. For example, when the sample is a whole blood sample, the electrode can perform a hematocrit measurement of the sample, which can be critical in determining the precise concentration of the analyte to be detected. Conductivity and/or impedance measurements can be determined depending on the sample being studied. Thus, the cassette of the present invention can detect whether an analyte is present in the sample, not only by binding/reaction with any analyte, but also electrical measurement of the sample. The electrodes can also be used to confirm that removal of the sample from the detection zone by the gas exiting the gas-filled chamber has occurred correctly because the conductivity detected when the liquid is present or absent can vary significantly. Electrodes may also be provided to the gas filled chamber to signal the compression of each of the chambers.

施加至盒之樣本可為任何合適液體樣本。它可為例如從受試者獲得之流體之樣本，諸如全血、血漿、唾液、***、汗、血清、月經、羊水、淚液、組織拭子、尿液、腦脊液、黏液樣本等。應瞭解本發明之檢定系統可應用於人類健康領域，包括較大及不斷增長的IVD市場(例如，癌症、心臟學、藥物濫用偵測及感染性疾病、包括細菌、真菌及病毒感染)。檢定亦可用於測試藥物及藥物作用。然而，系統亦可應用於環境背景，其中需要偵測例如毒性劑或傳染性病原體諸如細菌、真菌或病毒。因此，可獲取來自河流或湖泊或固體表面之拭子的樣本以便獲得提供至盒的液體樣本。檢定系統亦可用於獸醫應用。基本上，樣本可以液體形式提供或以液體形式呈現的任何檢定可用於本發明，例如，呼吸樣本可藉由向液體中吹氣來獲得並且該液體根據本發明來使用。亦可獲取表面之拭子並且安置在液體內以便提供液體樣本。 The sample applied to the cartridge can be any suitable liquid sample. It can be, for example, a sample of fluid obtained from a subject, such as whole blood, plasma, saliva, semen, sweat, serum, menses, amniotic fluid, tears, tissue swabs, urine, cerebrospinal fluid, mucus samples, and the like. It will be appreciated that the assay system of the present invention can be applied to the human health field, including the larger and growing market for IVD (eg, cancer, cardiology, drug abuse detection, and infectious diseases, including bacterial, fungal, and viral infections). Verification can also be used to test drugs and drug effects. However, the system can also be applied to environmental settings where it is desirable to detect, for example, toxic agents or infectious pathogens such as bacteria, fungi or viruses. Thus, a sample of a swab from a river or lake or solid surface can be obtained to obtain a liquid sample provided to the cartridge. The assay system can also be used for veterinary applications. Basically, any assay in which the sample may be provided in liquid form or presented in liquid form may be used in the present invention, for example, a breath sample may be obtained by insufflation into a liquid and the liquid is used in accordance with the present invention. A swab of the surface can also be obtained and placed in the liquid to provide a liquid sample.

樣本可例如包括直接從來源獲得之材料，諸如全血樣本，以及使用諸如過濾、沉澱、稀釋、蒸餾、混合、濃縮、鈍化干擾劑等之技術來預處理的材料。此等步驟可在樣本引入盒之前執行或可藉由盒本身來執 行。 The sample may, for example, include materials obtained directly from the source, such as whole blood samples, as well as materials pretreated using techniques such as filtration, precipitation, dilution, distillation, mixing, concentration, passivation of interfering agents, and the like. These steps can be performed before the sample is introduced into the cartridge or can be performed by the cartridge itself.

樣本可在盒***讀數器裝置之前或在盒***讀數器之後引入。在一些實施方式中，在施加樣本之前，盒***讀數器裝置中並且將力施加至氣體填充腔室以便從該/每個腔室排出氣體。此可有效地將盒準備好用於樣本施加。減少施加至該/每個腔室之力將氣體抽吸回到腔室中，進而將樣本抽吸至及沿著該/每個微流控通道。盒亦可被設計成使得樣本可最初經由毛細管作用來引入。以此方式，可提供如上所述之止擋特徵以便限制樣本進入該微流控通道中。由於將氣體從該/每個氣體填充腔室中排出或引入該/每個氣體填充腔室中而導致樣本之進一步傳送。為了樣本可最初經由毛細管作用來引入，需要存在於該微流控通道中之氣體藉由樣本來移位。此可經由從微流控通道退出至盒外部的閥等來達成。在一實施方式中，閥係單向閥，其被設計來僅允許氣體退出盒並且不允許氣體或液體引入盒中。 The sample can be introduced before the cartridge is inserted into the reader device or after the cartridge is inserted into the reader. In some embodiments, prior to applying the sample, the cartridge is inserted into the reader device and a force is applied to the gas-filled chamber to vent gas from the/each chamber. This effectively prepares the cartridge for sample application. Reducing the force applied to the/each chamber draws gas back into the chamber, thereby drawing the sample to and along the/each microfluidic channel. The cassette can also be designed such that the sample can be initially introduced via capillary action. In this manner, a stop feature as described above can be provided to limit the entry of sample into the microfluidic channel. Further transfer of the sample is caused by expelling gas from the/each gas-filled chamber or into the/each gas-filled chamber. In order for the sample to be initially introduced via capillary action, the gas present in the microfluidic channel needs to be displaced by the sample. This can be achieved via a valve or the like that exits from the microfluidic channel to the outside of the cartridge. In one embodiment, the valve train is designed to allow only gas to exit the cartridge and does not allow gas or liquid to be introduced into the cartridge.

閥可為例如小孔或縫隙，其與疏水性止擋特徵相鄰或緊鄰來定位，該止擋特徵被設計來防止僅藉由毛細管作用使樣本在該/每個微流控通道內進一步傳送。每個閥可經由比該/每個微流控通道本身更小尺寸之通道來與該/每個微流控通道流體連通(諸如該/每個微流控通道之寬度之小於50%、25%或20%)。在使用中，在反應過程發生之後，當樣本沿著該/每個微流控通道從偵測區移除時，樣本有利地朝向樣本輸入埠及/或可能存在之流體廢物吸收器來引導，而不是朝向閥來引導，此歸因於該/每個微流控通道之尺寸大於將微流控通道連接至閥的通道。此外，在最初樣本施加後，少量樣本可填充較小尺寸之通道並且充當在樣本施加之後閥與微流控通道之間之進一步流體流動的屏障。不受理論約束，預期屏障藉由與該較大主 要微流控通道相比，較小通道之相對更高毛細管現象造成。在毛細管填充之後，少量樣本可保留在較小通道中並且有效地將閥封堵。以此方式，閥僅在最初毛細管填充期間具有作用，其後在盒內之液體傳送藉由將氣體抽吸至該/每個腔室中或從該/每個腔室中排出來實現或控制。 The valve may be, for example, a small hole or slit that is positioned adjacent or in close proximity to the hydrophobic stop feature, the stop feature being designed to prevent further transfer of the sample within the/each microfluidic channel by capillary action only . Each valve may be in fluid communication with the/each microfluidic channel via a channel that is smaller than the/each microfluidic channel itself (such as less than 50% of the width of the/each microfluidic channel, 25 % or 20%). In use, after the reaction process occurs, as the sample is removed from the detection zone along the/each microfluidic channel, the sample is advantageously directed toward the sample input port and/or the fluid waste absorber that may be present, Rather than being directed toward the valve, this is due to the size of the/each microfluidic channel being larger than the channel connecting the microfluidic channel to the valve. Furthermore, after the initial sample application, a small sample can fill a smaller sized channel and act as a barrier to further fluid flow between the valve and the microfluidic channel after sample application. Without being bound by theory, it is expected that the barrier will result from a relatively higher capillary phenomenon of the smaller channel than the larger primary microfluidic channel. After capillary filling, a small amount of sample can remain in the smaller channel and effectively seal the valve. In this way, the valve only has an effect during the initial capillary filling, after which liquid transfer within the cartridge is achieved or controlled by pumping gas into or out of the chamber/each chamber. .

在另一態樣中，提供根據本發明使用之閥系統，該閥系統包含：根據本發明之檢定系統之頂部或底部表面中之通氣孔或縫隙開口；及通向檢定系統之該微流控通道的較小尺寸之微流控通道，該較小尺寸之微流控通道與檢定系統之通氣孔或縫隙開口及該微流控通道流體連通。 In another aspect, a valve system for use in accordance with the present invention is provided, the valve system comprising: a vent or slit opening in a top or bottom surface of the assay system in accordance with the present invention; and the microfluidic flow to the assay system A smaller sized microfluidic channel of the channel, the smaller sized microfluidic channel being in fluid communication with the vent or slit opening of the assay system and the microfluidic channel.

便利地，閥系統被定位成與該微流控通道之毛細管止擋物相鄰，以使得在樣本引入檢定系統後，樣本藉由毛細管作用來填充直至毛細管止擋物並且樣本之一部分亦至少部分地填充較小尺寸之微通道。至少部分地填充較小尺寸之微通道的樣本之一部分充當沿著較小尺寸之微通道之進一步流體傳送及經由通氣孔或縫隙之流體輸出的屏障。 Conveniently, the valve system is positioned adjacent to the capillary stop of the microfluidic channel such that after the sample is introduced into the assay system, the sample is filled by capillary action until the capillary stop and one portion of the sample is at least partially The ground fills the smaller size microchannel. One portion of the sample that at least partially fills the smaller sized microchannel acts as a barrier for further fluid transfer along the smaller sized microchannel and fluid output through the vent or slit.

合意地，本發明之盒可被設計來對於單一液體樣本進行複數個檢定(相同檢定之重複及/或不同檢定)。盒及相關聯通道之尺寸使得所有此等檢定理想地從液體樣本來執行，諸如可藉由手指穿刺來獲得的血液樣本，少於100ml、50ml，諸如少於40ml、30ml或甚至20ml或更少。以此方式，可在使用少於10ml，諸如少於7ml、5ml或甚至2ml或更少之液體樣本諸如血液的盒之單一通道中進行檢定。此大大少於在醫院中使用較大檯面 分析器或其他已知照護點平臺來執行分析所需要的樣本。 Desirably, the cartridge of the present invention can be designed to perform a plurality of assays for a single liquid sample (repetition of the same assay and/or different assays). The size of the cartridge and associated channel is such that all such assays are ideally performed from a liquid sample, such as a blood sample obtainable by finger puncture, less than 100 ml, 50 ml, such as less than 40 ml, 30 ml or even 20 ml or less. . In this way, the assay can be performed in a single channel using a cartridge of less than 10 ml, such as less than 7 ml, 5 ml or even 2 ml or less of a liquid sample such as blood. This is much less than the sample required to perform an analysis using a large benchtop analyzer or other known point-of-care platform in a hospital.

將要偵測之分析物可為任何所需分析物並且可包括蛋白質、肽、抗體、核酸、微生物(諸如細菌、真菌及病毒)、化學劑、生物化學劑、毒素、藥品、酶、代謝物、細胞部分、抗原等。舉例而言，本發明系統可適於偵測任何類型之分析物，該分析物可結合合適結合劑，或與合適試劑反應，該試劑之產物能夠偵測並且視需要藉由合適結合劑來結合。結合劑可為能夠特異性結合至將要偵測之分析物或反應產物的任何合適劑。舉例而言，若分析物係蛋白質或肽，結合劑可為能夠特異性結合至蛋白質/肽的受體或抗體。相反地，抗體可藉由該抗體被設計來特異性結合之蛋白質/肽來結合。核酸可藉由能夠特異性雜交至分析物核酸的其他核酸來結合。微生物可藉由特異性結合至微生物之表面上之蛋白質的抗體來結合。化學劑、毒素、藥品、代謝物可藉由化學部分來結合，該等化學部分能夠經由合適鍵結反應或親和力來反應或結合至前述化學分析物。許多類型之結合技術為熟習此項技術者熟知的。 The analyte to be detected can be any desired analyte and can include proteins, peptides, antibodies, nucleic acids, microorganisms (such as bacteria, fungi, and viruses), chemicals, biochemicals, toxins, drugs, enzymes, metabolites, Cell parts, antigens, etc. For example, the system of the present invention can be adapted to detect any type of analyte that can be combined with a suitable binding agent or reacted with a suitable reagent that is capable of detecting and, if desired, bound by a suitable binding agent. . The binding agent can be any suitable agent capable of specifically binding to the analyte or reaction product to be detected. For example, if the analyte is a protein or peptide, the binding agent can be a receptor or antibody capable of specifically binding to the protein/peptide. Conversely, an antibody can be bound by a protein/peptide that the antibody is designed to specifically bind to. Nucleic acids can be combined by other nucleic acids that are capable of specifically hybridizing to the analyte nucleic acid. Microorganisms can be bound by antibodies that specifically bind to proteins on the surface of the microorganism. Chemical agents, toxins, drugs, metabolites can be combined by chemical moieties that are capable of reacting or binding to the aforementioned chemical analytes via a suitable bonding reaction or affinity. Many types of bonding techniques are well known to those skilled in the art.

此外，該試劑可為酶或酶受質。舉例而言，分析物諸如葡萄糖可經由較好描述之酶促方法來偵測，因為在酶與葡萄糖反應之後所形成的反應產物可藉由使用審閱本說明書之熟習此項技術者已知之電化學或光學偵測技術來偵測。此等量測可作為獨立量測或與樣本中的將要偵測的其他分析物組合來進行。 In addition, the reagent can be an enzyme or an enzyme substrate. For example, an analyte such as glucose can be detected by a well-described enzymatic method, as the reaction product formed after the reaction of the enzyme with glucose can be electrochemically known to those skilled in the art by reviewing this specification. Or optical detection technology to detect. These measurements can be performed as separate measurements or in combination with other analytes in the sample to be detected.

應瞭解本文提到分析物/分析物結合劑複合物包括以下複合物，其中分析物未從其在液體樣本中發現之形式加以修飾，或其中分析物經由與另一個試劑反應來修飾並且由此可被視為分析物反應產物。 It is to be understood that the analyte/analyte binder complexes referred to herein include complexes in which the analyte is not modified from its form found in a liquid sample, or wherein the analyte is modified by reaction with another reagent and thereby Can be considered as an analyte reaction product.

結合劑可本身直接地或藉由合適鍵結至壁或表面來間接地附接至盒內之該微流控通道之壁或表面，例如，經由物理吸附、共價化學偶合、非共價化學鍵結(例如，生物素-抗生物素蛋白)或上述任何組合。在一個較佳實施方式中，結合劑呈包含結合部分的磁性或順磁粒子形式，並且結合部分可直接地或例如藉由非共價化學鍵結(例如，生物素-抗生物素蛋白)來間接地結合至粒子之表面。額外實施方式亦可包括物理吸附、共價化學偶合、非共價化學鍵結(例如，生物素-抗生物素蛋白)或其任何組合來結合至磁性劑，諸如磁性粒子之表面。經官能化以包含與其結合之結合劑的磁性劑/粒子可簡單置放在盒之該微流控通道內，以使得在將樣本施加至盒並且抽吸至並且沿著該/每個通道之後，官能化磁性劑/粒子藉由液體樣本來再懸浮，由此與樣本中之任何分析物接觸。結合及/或其他試劑之一或多個置放區域可經由先前描述之技術在置放區域之一或兩端處使用疏水性止擋物或其他特徵來專門界定，以便視需要將此一或多個區域與偵測區域/區分離。在適當情況下，此可確保不會由於在量測/偵測區域/區中之試劑組分(例如，螢光膠乳粒子)乾掉而導致獲得較高背景讀數。 The binding agent can be indirectly attached to the wall or surface of the microfluidic channel within the cartridge either directly or by suitable bonding to a wall or surface, for example, via physical adsorption, covalent chemical coupling, non-covalent chemical bonding A knot (eg, biotin-avidin) or any combination of the above. In a preferred embodiment, the binding agent is in the form of a magnetic or paramagnetic particle comprising a binding moiety, and the binding moiety can be indirectly or indirectly, for example, by non-covalent chemical bonding (eg, biotin-avidin) The ground is bonded to the surface of the particle. Additional embodiments may also include physical adsorption, covalent chemical coupling, non-covalent chemical bonding (eg, biotin-avidin), or any combination thereof, for binding to a magnetic agent, such as the surface of a magnetic particle. The magnetic agent/particles functionalized to include a binding agent bound thereto can be simply placed in the microfluidic channel of the cartridge such that after the sample is applied to the cartridge and aspirated to and along the/each channel The functionalized magnetic agent/particle is resuspended by the liquid sample, thereby contacting any analyte in the sample. One or more of the binding and/or other reagents may be specifically defined using hydrophobic stops or other features at one or both of the placement regions via techniques previously described, such that one or more Multiple areas are separated from the detection area/area. Where appropriate, this ensures that a higher background reading is not obtained due to the drying of reagent components (e.g., fluorescent latex particles) in the measurement/detection zone/area.

如上所述，除了黏合劑以外，盒可及/或替代地包含置放在該微流控通道內之一或多個試劑，該等試劑可促進偵測分析物或捕獲分析物。舉例而言，該一或多個試劑可包括適於特異性結合至分析物，由此促進其偵測的標記物，或與分析物反應以便產生分析物反應產物的酶。因此，根據本發明，本文所述檢定可用於偵測分析物或其分析物反應產物。 As noted above, in addition to the binder, the cartridge can and/or alternatively comprise one or more reagents disposed within the microfluidic channel that facilitate detection of the analyte or capture of the analyte. For example, the one or more reagents can include a label suitable for specific binding to the analyte, thereby facilitating its detection, or an enzyme that reacts with the analyte to produce an analyte reaction product. Thus, in accordance with the present invention, the assays described herein can be used to detect analytes or their analyte reaction products.

結合分析物可直接偵測，只要結合分析物能夠產生可偵測信號，或在結合分析物後，可發生反應，以便產生反應產物並且反應產物可 偵測到。然而，在一個較佳實施方式中，結合分析物與能夠結合至結合分析物的標記物接觸並且隨後偵測標記物/結合劑/分析物複合物。標記物可本身結合至另一個結合部分，該結合部分亦能夠特異性結合至結合劑/分析物複合物。通常標記物能夠結合至第一結合劑結合之分析物之不同部分，或能夠結合至結合劑/分析物複合物之區域，該區域僅在產生此複合物時形成。 The binding analyte can be detected directly, as long as the binding analyte is capable of generating a detectable signal, or after binding the analyte, a reaction can occur to produce a reaction product and the reaction product can be detected. However, in a preferred embodiment, the binding analyte is contacted with a label capable of binding to the binding analyte and the marker/binding agent/analyte complex is subsequently detected. The label may itself bind to another binding moiety that is also capable of specifically binding to the binding agent/analyte complex. Typically the label is capable of binding to a different portion of the analyte bound by the first binder, or is capable of binding to a region of the binder/analyte complex that is only formed upon production of the complex.

經由將氣體抽吸回到流體填充腔室，由此將流體樣本在該/每個氣體填充腔室之方向上進一步沿著微流控通道抽吸，可將結合分析物傳送至微流控通道之不同區域內之標記物。 The bound analyte can be delivered to the microfluidic channel by pumping gas back into the fluid-filled chamber, thereby drawing the fluid sample further along the microfluidic channel in the direction of the/each gas-filled chamber Markers in different areas.

合意地，結合劑及任何偵測劑/標記物在置放在盒之微流控通道中時呈乾燥狀態，以使得其能夠長期儲存，並且在液體樣本流動至及沿著微流控通道時藉由液體樣本來重構。 Desirably, the binding agent and any detection agent/label are in a dry state when placed in the microfluidic channel of the cartridge to enable long-term storage and when the liquid sample flows to and along the microfluidic channel Reconstituted by a liquid sample.

在一實施方式中，被設計來促進偵測分析物之結合及/或偵測劑/標記物最初定位在第一止擋特徵之下游(在引入之後，樣本流至盒中之方向上)。以此方式，在最初樣本施加及毛細管填充在盒內時，該結合劑及/或偵測劑最初不與樣本接觸。只在施加至該/每個氣體填充腔室之力減少並且氣體被抽吸回到氣體填充腔室中時，樣本進一步沿著該/每個微流控通道抽吸並且與結合劑及/或偵測劑接觸。 In one embodiment, it is designed to facilitate detection of binding of the analyte and/or the detection agent/label is initially positioned downstream of the first stop feature (after introduction, the sample flows into the direction of the cartridge). In this manner, the binding agent and/or detection agent is initially not in contact with the sample when the initial sample application and capillary filling are in the cartridge. The sample is further drawn along the/each microfluidic channel and with the binding agent and/or only when the force applied to the/each gas-filled chamber is reduced and the gas is drawn back into the gas-filled chamber. Detection agent contact.

在一實施方式中，樣本沿著微流控通道之傳送可在複數個級段中發生。舉例而言，在最初樣本施加及毛細管填充之後，藉由施加至該/每個氣體腔室之力之受控減少並且以受控及精確方式使氣體被抽吸回到該/每個氣體填充腔室中，樣本可沿著該/每個微流控通道之第一部分抽吸。該/每個微流控通道之第一部分可包含例如該結合劑。因此將流體樣本引入第 一部分中允許結合劑與可存在於液體樣本中之任何分析物反應。其後，藉由施加至該/每個氣體腔室之力之進一步受控減少，以使得更多氣體被抽吸至該/每個氣體填充腔室中，進而將樣本及結合劑抽吸至該/每個通道之第二部分中，樣本及結合劑可被抽吸至該/每個微流控通道之第二部分。另一個試劑或標記物例如可存在於第二部分中並且樣本及結合劑與其接觸。以此方式，可容易地實現關於特定檢定之多個分離步驟或級段並且每個步驟/級段可需要彼此不同的時間週期。應瞭解可容易地設想兩個以上級段，諸如三個、四個或更級段，並且每個級段藉由施加至該/每個氣體填充腔室之力之進一步受控減少來實現。有利地，每個氣體腔室可獨立地控制。以此方式，亦可使用本發明之單一盒來進行複數個不同類型之檢定。以此方式，每個分離通道設置有用於進行一或多個特定檢定的必需試劑並且讀數器經程式規劃以針對每個特定檢定來實現所需數目之氣體腔室壓縮/減壓步驟。因此，本發明之盒及相關聯讀數器能夠實質上同時進行多個分離及不同檢定，該等檢定可需要不同的試劑、反應時間週期、步驟數目等。 In an embodiment, the transfer of the sample along the microfluidic channel can occur in a plurality of stages. For example, after initial sample application and capillary filling, a controlled reduction in the force applied to the/each gas chamber and the gas being pumped back to the/each gas fill in a controlled and precise manner In the chamber, a sample can be drawn along the first portion of the/each microfluidic channel. The first portion of the/each microfluidic channel can comprise, for example, the binding agent. Thus introducing a fluid sample into the first portion allows the binding agent to react with any analyte that may be present in the liquid sample. Thereafter, a further controlled reduction in the force applied to the/each gas chamber causes more gas to be drawn into the/each gas-filled chamber, thereby pumping the sample and the binding agent to In the second portion of the/each channel, the sample and binding agent can be pumped to the second portion of the/each microfluidic channel. Another reagent or label can be present, for example, in the second portion and the sample and binding agent are in contact therewith. In this way, multiple separation steps or stages with respect to a particular assay can be readily implemented and each step/stage can require different time periods from each other. It will be appreciated that more than two stages, such as three, four or more stages, can be readily envisioned, and each stage is achieved by a further controlled reduction in the force applied to the/each gas-filled chamber. Advantageously, each gas chamber can be independently controlled. In this manner, a single cassette of the present invention can also be used to perform a plurality of different types of assays. In this manner, each separation channel is provided with the necessary reagents for performing one or more specific assays and the reader is programmed to achieve the desired number of gas chamber compression/decompression steps for each particular assay. Thus, the cartridge of the present invention and associated readers are capable of performing multiple separations and different assays substantially simultaneously, which may require different reagents, reaction time periods, number of steps, and the like.

雖然以上描述論述以逐步方式來抽吸或推動液體樣本，應認識到由於力控制構件之可控制性質，可在微小或可變的程度上可逆地壓縮及減壓氣體填充腔室，以便諸如允許在任何時間點來推動及拉動液體樣本並且允許混合效應。因此，例如，當液體樣本被傳送至包括將要藉由液體樣本來重構之一或多個試劑的微流控通道區域時，在到達該一或多個試劑置放之區域後，藉由使用每個/該氣體填充腔室之較小壓縮/減壓，可來回地推動及拉動液體樣本歷時一段時間，以便促進該一或多個試劑在液體樣本內之重構及/或混合。 While the above description discusses pumping or pushing a liquid sample in a stepwise manner, it will be appreciated that due to the controllable nature of the force control member, the gas can be reversibly compressed and decompressed to a small or variable extent, such as to allow for Push and pull the liquid sample at any point in time and allow mixing effects. Thus, for example, when a liquid sample is delivered to a microfluidic channel region that includes one or more reagents to be reconstituted by the liquid sample, after reaching the region in which the one or more reagents are placed, by using The smaller compression/decompression of each/the gas-filled chamber can push and pull the liquid sample back and forth for a period of time to facilitate reconstitution and/or mixing of the one or more reagents within the liquid sample.

本發明之方法及檢定之必要控制及實行可藉由使用讀數器內之合適微處理裝置及相關聯軟體來促進。 The necessary control and implementation of the methods and assays of the present invention can be facilitated by the use of suitable microprocessing devices and associated software within the reader.

在另一實施方式中，在樣本與諸如磁性粒子之結合劑之間之最初結合階段之後，在樣本液體內形成之結合劑-分析物複合物可傳送至通道之下游區域，其中標記物以乾燥形式定位在微流控通道內。樣本液體將標記物再懸浮/再水化並且允許標記物結合至結合劑-分析物複合物。液體樣本及任何重構材料之此傳送係歸因於每個/該氣體填充腔室上之減壓，從而將氣體抽吸回到每個/該氣體填充腔室中。將氣體/空氣抽吸回到每個/該氣體填充腔室中導致真空效應，其用來將液體樣本沿著該/每個微流控通道抽吸。此方法可允許所置放試劑之再水化及試劑分散之均勻性的更大控制。 In another embodiment, the binder-analyte complex formed within the sample liquid can be delivered to a downstream region of the channel after the initial binding phase between the sample and a binding agent such as magnetic particles, wherein the label is dried The form is positioned within the microfluidic channel. The sample liquid resuspends/rehydrates the label and allows the label to bind to the binder-analyte complex. This transfer of liquid sample and any reconstituted material is attributed to the reduced pressure on each/the gas-filled chamber, thereby drawing gas back into each/the gas-filled chamber. Pumping gas/air back into each/the gas-filled chamber results in a vacuum effect that is used to draw a liquid sample along the/each microfluidic channel. This method allows for greater control of the rehydration of the reagents placed and the uniformity of reagent dispersion.

在另一實施方式中，結合劑及標記物置放在該/每個微流控通道之同一區域中。樣本實質上同時將此等試劑再水化，使得結合及標記反應同時發生。在此實施方式中，所有試劑可接觸樣本，然後，經由施加磁鐵/電磁鐵，讀數器將磁性粒子-分析物-標記物複合物積聚在偵測區域內。不同於其他先前技術裝置，磁鐵/磁力可被設計成僅將磁性粒子積聚或集中在偵測區內。因此，磁鐵/磁力可不用來將磁性粒子沿著該/每個微流控通道縱向地抽吸或移動，而是實情為將任何複合物集中並保持在偵測區之區域中。在一實施方式中，磁性粒子可最初置放在微流控通道內的與磁力施加之位置相反的位置處。舉例而言，磁性粒子可在或沿著通道之底部置放並且磁鐵或磁力接觸/施加至盒之頂部表面。以此方式，在施加磁力時，磁性粒子被側向地(或垂直於通道內之液體樣本之流動)吸引穿過通道。預期主動地吸引磁性粒子穿過液體樣本之過程增加可在官能化磁性粒子與可存在於 液體樣本中之分析物之間發生的可能捕獲事件之數目。 In another embodiment, the binding agent and label are placed in the same region of the/each microfluidic channel. The sample is substantially rehydrated at the same time such that the binding and labeling reactions occur simultaneously. In this embodiment, all of the reagents can contact the sample, and then the magnetic particle-analyte-label complex is accumulated in the detection zone via the application of a magnet/electromagnet. Unlike other prior art devices, the magnet/magnetic force can be designed to concentrate or concentrate only magnetic particles within the detection zone. Thus, the magnet/magnetic force may not be used to draw or move the magnetic particles longitudinally along the/each microfluidic channel, but rather to concentrate and hold any complex in the region of the detection zone. In one embodiment, the magnetic particles may be initially placed at a location within the microfluidic channel that is opposite the location of the magnetic application. For example, the magnetic particles can be placed at or along the bottom of the channel and magnetized or magnetically contacted/applied to the top surface of the cartridge. In this way, upon application of a magnetic force, the magnetic particles are attracted laterally (or perpendicular to the flow of the liquid sample within the channel) through the channel. The process of actively attracting magnetic particles through the liquid sample is expected to increase the number of possible capture events that can occur between the functionalized magnetic particles and the analytes that may be present in the liquid sample.

在一實施方式中，提供電磁鐵，其被定位成與一旦正確***讀數器內之盒之偵測區一致。粒子-分析物-標記物複合物可藉由受控流體運動來抽吸至偵測區並且僅一旦處在偵測區中，即施加電磁力。另外，電磁鐵可經調適以便提供偵測區內之聚焦磁場。此可用來將粒子-分析物-標記物複合物集中在偵測區之規定部分，而不是遍及整個偵測區。替代磁鐵存在於讀數器中，可將磁鐵或合適電磁場產生電路提供在盒本身內。舉例而言，電磁場產生電路可相鄰於偵測區定位並且包括用於接觸讀數器內之對應連接器的電氣連接器等。一旦連接在一起，讀數器能夠提供產生電磁力之必需電信號。 In one embodiment, an electromagnet is provided that is positioned to coincide with the detection zone of the cartridge once properly inserted into the reader. The particle-analyte-label complex can be pumped to the detection zone by controlled fluid motion and applied electromagnetic force only once in the detection zone. Additionally, the electromagnet can be adapted to provide a focused magnetic field within the detection zone. This can be used to concentrate the particle-analyte-label complex in a defined portion of the detection zone rather than throughout the detection zone. Instead of a magnet present in the reader, a magnet or suitable electromagnetic field generating circuit can be provided within the cartridge itself. For example, the electromagnetic field generating circuit can be positioned adjacent to the detection zone and include an electrical connector or the like for contacting a corresponding connector within the reader. Once connected together, the reader can provide the necessary electrical signals to generate electromagnetic force.

為了促進偵測結合分析物，可需要從將要偵測結合分析物的該/每個偵測區域中移除廢液樣本。在需要時，本發明達成此舉，方法係使用存在於該/每個氣體填充腔室中之氣體來移除/推動反應或廢液樣本遠離該/每個偵測區微流控通道，並且朝向及進入可能存在之廢物腔室。然後，結合分析物，諸如磁性粒子-分析物-標記物複合物可在實質上無液體或實質上氣體環境中偵測及/或定量。應瞭解結合分析物可仍然為「濕的」，亦即，可存在塗布、包圍或以其他方式與結合分析物締合之一些殘留液體，但是如審閱本說明書之熟習此項技術者所理解的結合分析物不以整體液體形式存在。舉例而言，雖然結合分析物不以整體液體形式存在，但是結合分析物可在偵測期間保持水化(例如，其不被視為處於「乾燥」狀態下)。 To facilitate detection of bound analytes, it may be desirable to remove the waste sample from the/each detection zone where the bound analyte will be detected. When desired, the present invention achieves this by using a gas present in the/each gas-filled chamber to remove/push the reaction or waste sample away from the/each detection zone microfluidic channel, and Orient and enter a waste chamber that may be present. The bound analyte, such as a magnetic particle-analyte-label complex, can then be detected and/or quantified in a substantially liquid-free or substantially gaseous environment. It will be appreciated that the bound analyte may still be "wet", i.e., there may be some residual liquid that coats, surrounds, or otherwise associates with the bound analyte, but as understood by those skilled in the art in reviewing this specification. The bound analyte does not exist as a unitary liquid. For example, although the bound analyte does not exist as a unitary liquid, the bound analyte may remain hydrated during detection (eg, it is not considered to be in a "dry" state).

有利地，經由謹慎控制氣體進入及離開該/每個氣體腔室之運動，本發明能夠精確地控制沿著每個通道在任一方向上之液體運動速 率。舉例而言，可需要置放在該/每個通道內之乾燥試劑之重構快速地發生，但是在進行任何必要反應之後的液體樣本及未結合材料之移除相對緩慢地發生。因此，讀數器及相關聯力控制構件能夠變化或改變排出/進入該/每個氣體腔室之氣體之速度，從而對於該/每個通道中之液體運動之速度/速率具有對應作用。不同檢定可需要不同重構及/或液體移除速度並且此亦可藉由力控制構件組合相關聯程式規劃或軟體來獨立地控制。 Advantageously, the present invention is capable of accurately controlling the rate of liquid movement in either direction along each channel by carefully controlling the movement of gas into and out of the/each gas chamber. For example, reconstitution of the dried reagents placed in the/each channel may occur rapidly, but removal of the liquid sample and unbound material after any necessary reactions occurs relatively slowly. Thus, the reader and associated force control member can vary or vary the velocity of the gas exiting/entering the/each gas chamber, thereby having a corresponding effect on the speed/rate of liquid motion in the/each channel. Different assays may require different reconfiguration and/or liquid removal velocities and this may also be independently controlled by the force control component combination associated program plan or software.

此外，經由精細控制力控制構件，可謹慎控制非常小體積之氣體排出/進入該/每個氣體腔室，伴以液體樣本之對應較小運動。舉例而言，排出或引入該/每個氣體填充腔室之氣體之體積可為小於或等於500nl，諸如小於或等於200nl、100nl，或甚至50nl、25nl或15nl、10nl或甚至更少之增量。此較小體積之氣體運動導致該/每個通道中之液體之非常小對應線性運動。在偵測在實質上無液體環境中執行的本發明之實施方式中，發明人觀察到可使用此等非常小體積之氣體來將液體樣本及/或未捕獲材料恰好從偵測區或甚至其一部分中移除，並且由此在整體液體及未捕獲材料藉由氣體移除之實質上無液體環境中提供所捕獲的分析物或分析物反應產物。此完全不同於此項技術中之所謂習知洗滌步驟，其在執行偵測步驟之前使用較大體積之流體，通常液體，來清洗樣本偵測區/結合分析物等。事實上，在本發明中使用之空氣可不被視為清洗液，而是實情為僅在其內移除液體樣本及未捕獲材料。因此，當液體樣本及/或未捕獲材料需要從偵測區移除時，本發明能夠使用在體積上與發生偵測之偵測區或其一部分的體積實質上相等(或極輕微地較大，例如，15nl、25nl、50nl、100nl或200nl)的氣體體積，因為此足以將液體樣本從偵測區或部分移除，在實質上無液體環境中 留下分析物或分析物反應產物。在習知洗滌步驟中，總體上需要與樣本體積相比許多體積之清洗液。 Furthermore, via the fine control force control member, it is possible to carefully control the discharge/into the gas chamber of a very small volume with a correspondingly small movement of the liquid sample. For example, the volume of gas exiting or introducing the/each gas-filled chamber may be less than or equal to 500 nl, such as less than or equal to 200 nl, 100 nl, or even 50 nl, 25 nl or 15 nl, 10 nl or even less increments. . This smaller volume of gas movement results in a very small corresponding linear motion of the liquid in the/each channel. In detecting embodiments of the invention performed in a substantially liquid-free environment, the inventors have observed that such very small volumes of gas can be used to bring the liquid sample and/or the uncaptured material from the detection zone or even its The portion is removed and thereby the captured analyte or analyte reaction product is provided in a substantially liquid-free environment in which the bulk liquid and the uncaptured material are removed by gas. This is completely different from the so-called conventional washing step in the art, which uses a larger volume of fluid, typically a liquid, to clean the sample detection zone/bound analyte, etc., prior to performing the detection step. In fact, the air used in the present invention may not be regarded as a cleaning liquid, but rather the liquid sample and the uncaptured material are only removed therein. Therefore, when the liquid sample and/or the uncaptured material need to be removed from the detection zone, the present invention can be used to be substantially equal in volume (or slightly slightly larger) than the volume of the detection zone or portion thereof in which the detection occurs. For example, a volume of gas of 15nl, 25nl, 50nl, 100nl or 200nl), as this is sufficient to remove the liquid sample from the detection zone or portion, leaving the analyte or analyte reaction product in a substantially liquid-free environment. In conventional washing steps, a large volume of cleaning fluid is generally required compared to the sample volume.

此外，相對而言，在需要對液體樣本傳送至該/每個通道中及/或液體樣本從每個/該偵測區中移除進行控制時，該/每個氣體腔室之僅較小比例，諸如小於50%、40%或25%之體積可為上述控制所需要。 In addition, the gas chamber is relatively small only when it is required to transfer the liquid sample to the/each channel and/or the liquid sample is removed from each/the detection zone for control. A ratio, such as less than 50%, 40% or 25% by volume, may be required for the above control.

每個盒可被設計來執行單一分析物偵測或多個分析物檢測。此外，每個盒可包含一個以上微流控通道系統，以使得可使用單一盒來執行一個以上檢定。亦可每個微流控通道執行一個以上檢定。以此方式，每個盒可執行來自單一液體樣本之許多重複及/或明顯不同檢定，因為該/每個氣體腔室獨立地可控制。 Each cassette can be designed to perform a single analyte detection or multiple analyte detection. In addition, each cartridge can contain more than one microfluidic channel system such that more than one assay can be performed using a single cartridge. More than one check can be performed per microfluidic channel. In this manner, each cartridge can perform many iterations and/or significantly different assays from a single liquid sample, as the/each gas chamber is independently controllable.

合意地，盒可容易地大規模生產。盒可以帶材形式提供，其中多個盒最初例如諸如經由有孔的密封件來連接在一起。以此方式，使用者可在使用之前容易地將盒從帶材中移除。 Desirably, the cartridge can be easily mass produced. The cartridge may be provided in the form of a strip, wherein the plurality of cartridges are initially joined together, for example, such as via a perforated seal. In this way, the user can easily remove the cassette from the strip prior to use.

一旦盒以樣本負載，任何所捕獲分析物可經由存在於讀數器裝置中之合適光學或其他構件來偵測。本發明提供此讀數器並且本發明之重要態樣係提供至少一個力控制構件，其存在於讀數器中並且被設計來控制施加至該一或多個氣體填充腔室之外表面之力，以便將氣體從該/每個氣體填充腔室排出/引入該/每個氣體填充腔室中。減少藉由力控制構件施加之力導致氣體被抽吸回到該/每個氣體填充腔室中。本發明之一個優勢在於盒本身可最初為「乾燥的」，在樣本施加之前，在盒內幾乎沒有整體液體。此不僅簡化盒本身之製造，而且改良保存限期並且允許本發明之許多盒在使用之前儲存在室溫下，並且盒內之化學或生物組分極少降級。 Once the cartridge is loaded with the sample, any captured analyte can be detected via suitable optical or other components present in the reader device. The present invention provides such a reader and an important aspect of the present invention provides at least one force control member present in the reader and designed to control the force applied to the outer surface of the one or more gas filled chambers so that Gas is discharged/introduced from the/each gas-filled chamber into the/each gas-filled chamber. Reducing the force applied by the force control member causes the gas to be drawn back into the/each gas filled chamber. One advantage of the present invention is that the cartridge itself can be initially "dry" with little overall liquid in the cartridge prior to application of the sample. This not only simplifies the manufacture of the cartridge itself, but also improves the shelf life and allows many of the cartridges of the present invention to be stored at room temperature prior to use, and the chemical or biological components within the cartridge are rarely degraded.

在另一態樣中，提供用於本發明之微流控系統的讀數器裝置，該讀數器裝置包含：力控制構件，其用於控制微流控系統之氣體填充腔室之壓縮或減壓；及偵測構件，其用於實現引入微流控盒中之液體樣本內之所需分析物，或其分析物反應產物的偵測；其中力控制構件包含壓電彎曲致動器，其被設計來直接地或經由致動器之位移來間接地實現氣體填充腔室之壓縮或減壓。 In another aspect, a reader device for use in a microfluidic system of the present invention is provided, the reader device comprising: a force control member for controlling compression or decompression of a gas filled chamber of a microfluidic system And a detecting member for effecting detection of a desired analyte introduced into the liquid sample in the microfluidic cartridge, or an analyte reaction product thereof; wherein the force control member comprises a piezoelectric bending actuator, which is It is designed to indirectly effect compression or decompression of the gas filled chamber, either directly or via displacement of the actuator.

皮埃爾居里在1883年發現了壓電效應。他注意到，某些材料，如石英晶體，在受到機械應力時會產生電壓。相反，當施加電壓時，彼等材料之形狀會變形。因此，它們可用作換能器，將電信號轉換成機械振動。 Pierre Curie discovered the piezoelectric effect in 1883. He noticed that certain materials, such as quartz crystals, generate voltage when subjected to mechanical stress. Conversely, when a voltage is applied, the shape of their materials will deform. Therefore, they can be used as transducers to convert electrical signals into mechanical vibrations.

各種材料具有壓電性質；最常用的是鋯酸鉛(PZT)。改變陶瓷之化學成分及製造過程可改變壓電彎曲機之性能。當PZT層連接至適當基片(例如薄金屬板)上時，PZT板之任何電活化導致該板相對於基板之平面運動，從而引起內部機械應力，導致類似於熱雙金屬之複合結構的彎曲運動。 Various materials have piezoelectric properties; the most commonly used is lead zirconate (PZT). Changing the chemical composition of the ceramic and the manufacturing process can change the performance of the piezoelectric bending machine. When the PZT layer is attached to a suitable substrate (e.g., a thin metal plate), any electrical activation of the PZT plate causes the plate to move relative to the plane of the substrate, causing internal mechanical stresses, resulting in bending of a composite structure similar to a hot bimetal. motion.

壓電彎曲機在此項技術中為熟知的。通常，壓電陶瓷晶體可在兩側用銀塗布並且膠合至黃銅、鎳合金或不銹鋼帶材。 Piezoelectric bending machines are well known in the art. Typically, piezoelectric ceramic crystals can be coated with silver on both sides and glued to brass, nickel alloy or stainless steel strips.

陶瓷可被組配成具有或不具有回饋。回饋可與外電路結合使用以便監測壓電彎曲機之操作並且調整輸入信號以保持一致輸出頻率。 The ceramics can be grouped with or without feedback. Feedback can be used in conjunction with an external circuit to monitor the operation of the piezoelectric bender and adjust the input signal to maintain a consistent output frequency.

彎曲機可以從PZT雙層或多層結構切割的各種幾何形狀來製成。本發明之壓電彎曲機可採用帶材彎曲機形式。對於帶材彎曲機，將 帶材之一端固定安裝，並且另一端自由地運動：對於此安裝，達成帶材彎曲機之最大位移並且位移、剛度及共振之指定資料係指此情境。位移視帶材之自由運動長度而定。提供彎曲機之總長度的通常大約5-10%用於安裝目的。安裝可藉由夾持或藉由使用黏著劑如環氧樹脂、氰基丙烯酸酯等來進行。 The bender can be made from a variety of geometries cut from PZT double or multi-layer structures. The piezoelectric bending machine of the present invention can be in the form of a strip bending machine. For strip bending machines, one end of the strip is fixedly mounted and the other end is free to move: for this installation, the maximum displacement of the strip bending machine and the specified information on displacement, stiffness and resonance are referred to in this context. The displacement depends on the free movement length of the strip. Typically about 5-10% of the total length of the bender is provided for installation purposes. Mounting can be carried out by clamping or by using an adhesive such as epoxy resin, cyanoacrylate or the like.

壓電彎曲機可最初偏置，或導致相關聯底座或指狀物與氣體填充腔室之外表面接觸。以此方式，可最初提供施加至氣體腔室之最大力，導致氣體從氣體腔室排出。在發出適當電信號後，可誘導壓電彎曲機彎曲遠離氣體腔室之外表面，導致減少施加至腔室之力並且導致將氣體抽吸至腔室中。因為每個氣體腔室與該/每個微流控通道流體連通，審閱本說明書之熟習此項技術者容易理解排出或抽吸至相應氣體腔室中之氣體如何導致相應微流控通道中之液體樣本之對應定向運動。 The piezoelectric bender can be initially biased or cause the associated base or finger to contact the outer surface of the gas filled chamber. In this way, the maximum force applied to the gas chamber can be initially provided, causing the gas to exit the gas chamber. Upon emitting an appropriate electrical signal, the piezoelectric bender can be induced to bend away from the outer surface of the gas chamber, resulting in reduced forces applied to the chamber and causing gas to be drawn into the chamber. Because each gas chamber is in fluid communication with the/each microfluidic channel, it is readily understood by those skilled in the art to understand how the gas being vented or pumped into the respective gas chamber results in a corresponding microfluidic channel. Corresponding directional movement of the liquid sample.

讀數器可包括盒***其中的接收埠。讀數器可經調適以便確保盒之正確***並且此可採用各種形式。舉例而言，盒可最初定位在進入讀數器之載體機構中，諸如可在電腦中發現的用於負載CD等之載體機構。或者，接收埠可設定大小以允許接收盒並且可在讀數器內發現內部止擋構件，盒一旦正確地***即鄰接該內部止擋構件。另外或替代地，在盒之表面上發現或切割之特徵可被設計來與在讀數器中發現之特徵共置並且僅一旦盒正確定位在讀數器中，盒能夠藉由讀數器來控制。可提供不同大小的接收埠，或單一接收埠適當地成形以接收不同大小的盒，該等盒被設計來執行例如特定數目之檢定。 The reader can include a receiving cassette into which the cartridge is inserted. The reader can be adapted to ensure proper insertion of the cartridge and this can take a variety of forms. For example, the cartridge can be initially positioned in a carrier mechanism that enters the reader, such as a carrier mechanism that can be found in a computer for loading a CD or the like. Alternatively, the receiving cassette can be sized to allow receipt of the cartridge and an internal stop member can be found within the reader, the cartridge abutting the internal stop member once properly inserted. Additionally or alternatively, features found or cut on the surface of the cartridge can be designed to coexist with features found in the reader and can be controlled by the reader once the cartridge is properly positioned in the reader. Receivers of different sizes may be provided, or a single receiver, suitably shaped to receive boxes of different sizes, which are designed to perform, for example, a particular number of checks.

讀數器可經由合適軟體被組配來執行各種不同類型的檢 定。可向使用者提供套組，其包含檢定盒及視需要樣本收集裝置。盒可包含條形碼或讀數器裝置能夠判定之其他表面特徵，其可用來通知讀數器***讀數器中之盒之類型以及由此進行何等一或多個檢定及由此讀數器應該如何運作及/或例如提供患者詳細資料。以此方式，可提供單一類型之讀數器，其能夠接收可進行不同檢定及/或組檢定的各種不同盒。 The readers can be assembled via suitable software to perform various different types of assays. A kit can be provided to the user that includes a assay cartridge and an optional sample collection device. The cartridge may contain a barcode or other surface feature that the reader device can determine which can be used to inform the reader of the type of cartridge inserted into the reader and the one or more assays thereby and how the reader should operate and/or For example, provide patient details. In this manner, a single type of reader can be provided that is capable of receiving a variety of different cartridges that can perform different assays and/or group assays.

在結合劑結合至磁性劑，諸如磁性珠粒之表面的實施方式中，應瞭解讀數器包含永磁鐵或電磁鐵。磁鐵被設計來緊密鄰近磁性劑，或誘導電磁鐵來施加磁場，以便將磁性粒子集中並保持在盒之該微流控通道之特定區域中。此區域可為偵測區域。在一實施方式中，使用僅一旦磁性粒子傳送至偵測區即加以接通的電磁鐵。藉由合適設計，亦可控制或聚焦電磁鐵之磁場以確保磁性粒子聚焦並保持在偵測區之較小區域中。此可用來將磁性粒子積聚至較小區域並且增加可偵測到之信號。 In embodiments where the binding agent is bonded to a magnetic agent, such as the surface of a magnetic bead, it should be understood that the reader comprises a permanent magnet or an electromagnet. The magnet is designed to be in close proximity to the magnetic agent or to induce an electromagnet to apply a magnetic field to concentrate and hold the magnetic particles in a particular region of the microfluidic channel of the cartridge. This area can be the detection area. In one embodiment, an electromagnet that is turned on only once the magnetic particles are delivered to the detection zone is used. With a suitable design, the magnetic field of the electromagnet can also be controlled or focused to ensure that the magnetic particles are focused and held in a small area of the detection zone. This can be used to accumulate magnetic particles into smaller areas and increase the detectable signal.

將磁性粒子集中至特定區域可用來促進任何所捕獲分析物之偵測及/或增加偵測之靈敏度。此外，經由磁場來保持粒子，其亦允許藉由從該/每個氣體填充腔室排出之氣體來將包圍結合分析物的不必要的/耗盡的流體樣本加以移除，從而保持捕獲分析物不含可存在於最初樣本中之潛在干擾劑/污染物。永磁鐵或電磁場可得以減少或增加，方法係諸如藉由移動永磁鐵更接近於或進一步遠離盒，或增加或降低所施加場之強度。此可用來允許磁性粒子「鬆弛」或在特定位置中變得不太集中，同時仍然在一定程度上藉由磁場來保持或不保持。此可促進對於粒子執行進一步反應，與在磁性粒子緊密地集中時相比，該等反應可更有效地進行。在某些應用中亦可較佳地在粒子不太「集中」或鬆弛時執行偵測。 Concentration of magnetic particles to specific regions can be used to facilitate detection of any captured analyte and/or increase sensitivity of detection. In addition, the particles are held by the magnetic field, which also allows the removal of unnecessary/depleted fluid samples surrounding the bound analyte by the gas exiting the chamber/each gas-filled chamber, thereby maintaining the captured analyte Contains no potential interfering agents/contaminants that may be present in the original sample. Permanent magnets or electromagnetic fields can be reduced or increased by, for example, moving the permanent magnet closer to or further away from the cartridge, or increasing or decreasing the strength of the applied field. This can be used to allow the magnetic particles to "relax" or become less concentrated in a particular location while still being held or not maintained to some extent by the magnetic field. This can promote the further reaction to the particles, which can be carried out more efficiently than when the magnetic particles are densely concentrated. In some applications it is also preferred to perform detection when the particles are less "concentrated" or slack.

在使用中，一旦磁場施加至樣本，磁鐵可用於保持任何結合劑。氣體可從該/每個氣體填充腔室中排出以便傳送液體樣本及存在於樣本中之任何未結合組分遠離該/每個偵測區域且/或允許其他試劑諸如偵測劑與所捕獲的分析物接觸。為了確保氣體之力不足以將磁性結合材料剝離，謹慎控制氣體運動速度及對應液體樣本及任何未結合組分移除係必不可少的。因此，可謹慎控制從該/每個氣體填充腔室排出之氣體之速度。在某些實施方式中，可需要在施加磁場/力之前抽吸液體樣本及試劑等經過偵測區。因此，磁性粒子之任何捕獲僅在一旦藉由從氣體腔室排出之氣體將液體樣本向後推動穿過偵測區時發生。 In use, a magnet can be used to hold any binding agent once a magnetic field is applied to the sample. Gas may be expelled from the/each gas-filled chamber to transport the liquid sample and any unbound components present in the sample away from the/each detection zone and/or allow other reagents such as detection agents and captured Analyte contact. In order to ensure that the force of the gas is insufficient to strip the magnetically bound material, careful control of the rate of gas movement and the corresponding liquid sample and any unbound component removal are essential. Therefore, the speed of the gas discharged from the/each gas-filled chamber can be carefully controlled. In some embodiments, it may be desirable to pump a liquid sample, reagent, etc. through the detection zone prior to applying the magnetic field/force. Thus, any capture of the magnetic particles occurs only once the liquid sample is pushed back through the detection zone by the gas exiting the gas chamber.

在另一實施方式中，磁性粒子可以結合試劑塗布，該結合試劑被設計來移除樣本中之干擾。磁性粒子結合存在於樣本中之此干擾物並且然後，磁性粒子可保持在與特定捕獲/偵測試劑及/或偵測區分開的特定位置中以允許反應進行並且在不存在特定干擾物的情況下量測。 In another embodiment, the magnetic particles can be coated with a binding agent designed to remove interference in the sample. The magnetic particles bind to the interferer present in the sample and then the magnetic particles can remain in a particular location separate from the particular capture/detection reagent and/or detection to allow the reaction to proceed and in the absence of a particular interferer The next measurement.

本發明之讀數器進一步包含用於偵測樣本盒內之任何所捕獲分析物的偵測構件。視特定檢定而定，偵測構件可為任何合適構件。舉例而言，偵測構件可為螢光計或分光光度計，其可用於偵測一旦適當地激發即來自加標記或未加標記結合分析物或反應產物之螢光信號。一旦合適波長之光用於激發分析物/產物，結合分析物/反應產物可自然地發螢光，或另一個標記物可用於個別地結合至結合分析物並且標記物藉由螢光構件來偵測。可使用之其他標記物及由此相應地適合之偵測構件包括放射性標記、磷光標記、膠態金屬粒子、生物發光標記、比色標記、電化學標記等。此外，如上所述，分析物或其反應產物，或結合分析物或反應產物本身可 使用諸如拉曼光譜法等之技術來直接偵測。在一些實施方式中，偵測構件被設計來光學偵測分析物或分析物反應產物，或所捕獲分析物/分析物反應產物及/或附接至任何前述部分的標記物。 The reader of the present invention further includes a detection member for detecting any captured analyte within the sample cartridge. Depending on the particular assay, the detection member can be any suitable member. For example, the detection component can be a fluorometer or a spectrophotometer that can be used to detect a fluorescent signal from a labeled or unlabeled binding analyte or reaction product once properly excited. Once the appropriate wavelength of light is used to excite the analyte/product, the bound analyte/reaction product can naturally fluoresce, or another marker can be used to individually bind to the bound analyte and the label is detected by the fluorescent member. Measurement. Other labels that can be used and thus correspondingly suitable detection members include radioactive labels, phosphorescent labels, colloidal metal particles, bioluminescent labels, colorimetric labels, electrochemical labels, and the like. Further, as described above, the analyte or its reaction product, or the bound analyte or the reaction product itself, can be directly detected using a technique such as Raman spectroscopy. In some embodiments, the detection member is designed to optically detect an analyte or analyte reaction product, or a captured analyte/analyte reaction product and/or a label attached to any of the foregoing portions.

可偵測標記可單獨使用，或結合微觀粒子或珠粒，諸如金屬氧化物、多醣或膠乳粒子來使用。許多類型之膠乳及其他粒子在此項技術中為已知的。 The detectable label can be used alone or in combination with microscopic particles or beads, such as metal oxides, polysaccharides or latex particles. Many types of latex and other particles are known in the art.

讀數器包含力控制構件，其包含以上論述之一或多個壓電彎曲機，該等彎曲機用於接觸盒之該/每個氣體填充腔室並且藉由增加或減少藉由彎曲機形成之彎曲來減少或增加施加至該/每個氣體填充腔室的力。當提供一個以上氣體填充腔室時，可提供用於每個氣體填充腔室的分離獨立控制壓電彎曲機。力控制構件可包括指狀物或底座，其被設計來接觸該/每個腔室之外表面並且向其施加力。以此方式，壓電彎曲機作用於指狀物或底座，以使得指狀物/底座作用於氣體填充腔室。在使用中，在力控制構件接觸該/每個腔室之外表面之前，腔室處在最大容積、氣體填充狀態下。在使該/每個氣體填充腔室之表面與所施加力接觸後，該/每個腔室內之氣體得以排出。增加所施加之力導致進一步氣體從該/每個腔室排出。相反地，藉由力控制構件來減少施加至該/每個氣體填充腔室之力導致氣體被抽吸回到該/每個腔室中。 The reader comprises a force control member comprising one or more of the piezoelectric bending machines discussed above for use in the/each gas-filled chamber of the contact cartridge and formed by a bending machine by increasing or decreasing Bending to reduce or increase the force applied to the/each gas filled chamber. When more than one gas fill chamber is provided, a separate, independently controlled piezoelectric bender for each gas filled chamber can be provided. The force control member can include a finger or a base that is designed to contact and apply a force to the outer surface of the/each chamber. In this manner, the piezoelectric bending machine acts on the fingers or base such that the fingers/bases act on the gas filled chamber. In use, the chamber is in a maximum volume, gas filled state before the force control member contacts the outer surface of the/each chamber. After the surface of the/each gas-filled chamber is brought into contact with the applied force, the gas in the chamber is discharged. Increasing the applied force causes further gas to escape from the/each chamber. Conversely, reducing the force applied to the/each gas-filled chamber by the force control member causes gas to be drawn back into the/each chamber.

指狀物/底座可被設計來接觸該/每個腔室之外表面之中心部分。通常，指狀物/底座可僅接觸該/每個氣體填充腔室之總外表面之一部分。舉例而言，在使用中，指狀物/底座可接觸盒之頂部表面並且設定大小來接觸10與50%之間的覆蓋氣體腔室之頂部表面區域。使用如所描述之壓 電彎曲機，將與盒之表面接觸的指狀物/底座升高及降低，或迫使其與氣體腔室之表面接觸及鬆弛。力控制構件之速度及彎曲度及由此產生之操作可謹慎控制以便能夠控制排出或抽吸至該/每個氣體填充腔室中之氣體之速度及量。 The fingers/bases can be designed to contact a central portion of the outer surface of the/each chamber. Typically, the fingers/base may only contact a portion of the total outer surface of the/each gas-filled chamber. For example, in use, the fingers/base can contact the top surface of the cartridge and be sized to contact between 10 and 50% of the top surface area of the blanket gas chamber. Using a piezoelectric bending machine as described, the fingers/bases in contact with the surface of the box are raised and lowered, or forced into contact with the surface of the gas chamber and relaxed. The speed and curvature of the force control member and the resulting operation can be carefully controlled to enable control of the velocity and amount of gas that is vented or pumped into the/each gas filled chamber.

讀數器可包括其他特徵，諸如加熱裝置以允許檢定在特定溫度下進行，並且包括合適電路及軟體以允許讀數器經程式規劃以執行一或多個不同檢定。 The reader may include other features, such as a heating device to allow the assay to be performed at a particular temperature, and includes suitable circuitry and software to allow the reader to be programmed to perform one or more different assays.

在另一態樣中，提供檢定系統，其包含自給式微流控系統及相關聯讀數器裝置，其中：自給式微流控系統包含：樣本輸入埠，其用於接收將要檢定之液體樣本，該樣本輸入埠連接至至少一個微流控通道，其中每個/該微流控通道包含置放在其中以用於進行檢定的一或多個試劑及偵測區，該偵測區用於偵測可存在於樣本中之任何分析物或分析物反應產物；及每個/該微流控通道與每個/該偵測區下游之可壓縮、氣體填充腔室流體連通，其中微流控系統由三個層形成，該等層層疊在一起以界定每個/該微流控通道及該氣體填充腔室，並且其中使該腔室壓縮或減壓導致將氣體從腔室中排出或抽吸至腔室中，進而導致該/每個微流控通道內之液體樣本之運動；及與微流控系統一起使用之讀數器裝置，該讀數器裝置包含：力控制構件，其用於控制微流控系統之氣體填充腔室之壓縮 或減壓；及偵測構件，其用於實現引入微流控盒中之液體樣本內之所需分析物，或其分析物反應產物的偵測；其中力控制構件包含壓電彎曲致動器，其被設計來直接地或經由致動器之位移來間接地將氣體填充腔室壓縮或減壓。 In another aspect, an assay system is provided comprising a self-contained microfluidic system and associated reader device, wherein: the self-contained microfluidic system comprises: a sample input port for receiving a liquid sample to be assayed, the sample The input port is coupled to the at least one microfluidic channel, wherein each/the microfluidic channel includes one or more reagents and detection zones disposed therein for performing the assay, the detection zone being used for detecting Any analyte or analyte reaction product present in the sample; and each/the microfluidic channel is in fluid communication with each compressible, gas-filled chamber downstream of the detection zone, wherein the microfluidic system is comprised of three Layer formation, the layers being stacked together to define each/the microfluidic channel and the gas-filled chamber, and wherein compressing or decompressing the chamber results in expelling or pumping gas from the chamber to the chamber a chamber, which in turn causes movement of the liquid sample within the/each microfluidic channel; and a reader device for use with the microfluidic system, the reader device comprising: a force control member for controlling microfluidic control System gas Compressing or decompressing the filling chamber; and detecting means for realizing the desired analyte introduced into the liquid sample in the microfluidic cartridge, or detecting the analyte reaction product; wherein the force controlling member comprises pressure An electric bending actuator designed to indirectly compress or depressurize the gas filled chamber, either directly or via displacement of the actuator.

在另一態樣中，提供對於液體樣本進行檢定之方法，該方法包含：a)將如本文描述之微流控系統提供至如本文描述之讀數器裝置；b)壓縮微流控系統之一/該氣體填充腔室，以便將氣體從該/每個氣體填充腔室排出；c)將液體樣本引入微流控系統並且允許藉由毛細管作用將樣本抽吸至該/每個微流控通道中，且/或部分地將該/每個氣體填充腔室減壓以使得氣體被抽吸至該/每個腔室中，從而導致液體樣本被抽吸至該/每個微流控通道中；d)使得一或多個試劑與存在於液體樣本中之任何分析物反應；e)視需要部分地進一步部分地將微流控系統之該/每個氣體填充腔室減壓，以使得液體樣本進一步沿著該/每個微流控通道朝向該/每個氣體填充腔室抽吸並且視需要使液體樣本與分析物結合劑及/或一或多個其他試劑接觸；f)視需要捕獲任何分析物或分析物反應產物並且壓縮該/每個氣體填充腔室，以使得從該/每個腔室排出之氣體導致液體樣本及未捕獲材料被推動遠離任何所捕獲分析物或分析物反應產物；及g)偵測任何分析物或分析物反應產物，或所捕獲的分析物或分析物反 應產物。 In another aspect, a method of assaying a liquid sample is provided, the method comprising: a) providing a microfluidic system as described herein to a reader device as described herein; b) one of a compressed microfluidic system / the gas fills the chamber to discharge gas from the / each gas filled chamber; c) introduces a liquid sample into the microfluidic system and allows the sample to be pumped to the/each microfluidic channel by capillary action And/or partially decompressing the/each gas-filled chamber to cause gas to be drawn into the/each chamber, thereby causing a liquid sample to be drawn into the/each microfluidic channel d) causing one or more reagents to react with any analyte present in the liquid sample; e) partially partially decompressing the gas perfusion chamber of the microfluidic system as needed to cause liquid The sample is further drawn toward the/each gas-filled chamber along the/each microfluidic channel and the liquid sample is contacted with the analyte binding agent and/or one or more other reagents as needed; f) captured as needed Any analyte or analyte reaction product and Shrinking/each gas fills the chamber such that gas exiting the chamber/each chamber causes the liquid sample and the uncaptured material to be pushed away from any captured analyte or analyte reaction product; and g) detecting any analysis The analyte or analyte reaction product, or the captured analyte or analyte reaction product.

應瞭解上述方法之步驟e)可作為單一或多個步驟來執行。因此，視將要執行之檢定而定，步驟e)可為單一步驟以使得施加至該/每個氣體填充腔室之力之減少係力之單一減少並且樣本被抽吸至該/每個微流控通道中之單一位置。或者，可存在多個步驟，諸如2、3或4個步驟，其中力之連續減少施加至該/每個腔室，導致樣本被抽吸至該/每個微流控通道內之許多連續位置，視執行力之減少的次數而定。因此，本發明允許進行檢定，其中需要單一步驟或多個步驟。 It should be understood that step e) of the above method can be performed as a single or multiple steps. Thus, depending on the assay to be performed, step e) can be a single step such that the force of the force applied to the/each gas-filled chamber is reduced by a single force and the sample is pumped to the/each microfluid Control a single location in the channel. Alternatively, there may be multiple steps, such as 2, 3 or 4 steps, wherein a continuous reduction in force is applied to the/each chamber, causing the sample to be drawn to many consecutive locations within the/each microfluidic channel , depending on the number of times the execution is reduced. Thus, the present invention allows for verification in which a single step or multiple steps are required.

施加至氣體填充腔室之力可藉由如在上文中論述之力控制構件來提供，該等力控制構件包含壓電彎曲機及視需要與其關聯之指狀物或底座。 The force applied to the gas-filled chamber can be provided by a force control member as discussed above, the force control member comprising a piezoelectric bender and a finger or base associated therewith as desired.

施加至該/每個氣體填充腔室之力之增加或減少之速率可改變以便增加或減少該/每個通道中之液體運動之速度。舉例而言，在需要時，在步驟e)中施加之力之減少可比在步驟f)中施加之力之增加速率更快。 The rate of increase or decrease in the force applied to the/each gas-filled chamber can be varied to increase or decrease the rate of liquid movement in the/each channel. For example, the reduction in the force applied in step e) can be faster than the rate of increase in the force applied in step f), if desired.

分析物/分析物結合劑複合物之捕獲可歸因於例如結合至微流控通道之表面的分析物結合劑，或由於具有磁性並且向所形成複合物施加磁力而捕獲。用於形成複合物之磁性粒子可最初置放在與磁鐵緊密接觸，或施加磁力的盒之表面相反的該微流控通道之表面上。其作用係垂直於液體穿過該通道之流動，磁性粒子被側向地抽吸穿過該微流控通道，從而增加及/或增強磁性粒子與分析物或分析物反應產物之接觸，從而增加檢定之靈敏度。 The capture of the analyte/analyte binder complex can be attributable to, for example, an analyte binding agent bound to the surface of the microfluidic channel, or captured due to its magnetic properties and the application of a magnetic force to the formed complex. The magnetic particles used to form the composite may be initially placed on the surface of the microfluidic channel that is in intimate contact with the magnet or opposite the surface of the magnetic field. The action is perpendicular to the flow of liquid through the channel, and the magnetic particles are drawn laterally through the microfluidic channel to increase and/or enhance contact of the magnetic particles with the analyte or analyte reaction product, thereby increasing Sensitivity of the test.

可使用單一盒來執行上述方法之一個以上實施方式。因此， 例如，包括上述步驟f)之方法可在本發明之盒內之一個通道中執行並且不包括步驟f)之方法可在單獨通道上執行。另外或替代地，步驟e)可在前述通道及/或額外通道上單獨或多次執行。以此方式，多個不同類型之檢定可使用包含複數個檢定通道之單一盒來進行。 One or more embodiments of the above methods can be performed using a single cartridge. Thus, for example, the method comprising the above step f) can be performed in one of the channels within the cartridge of the present invention and the method not including the step f) can be performed on a separate channel. Additionally or alternatively, step e) may be performed alone or multiple times on the aforementioned channels and/or additional channels. In this manner, multiple different types of assays can be performed using a single cartridge containing a plurality of assay channels.

本發明進一步基於一種檢定系統之開發，該檢定系統包含能夠對於單一樣本進行多個不同檢定的拋棄式微流控盒及相關聯讀數器，該讀數器能夠偵測及/或判定來自單一樣本之複數個分析物之水準並且向使用者提供輸出。本發明亦允許藉由讀數器來接收各種拋棄式盒，該各種拋棄式盒中之每一者能夠執行一組獨特的不同檢定。以此方式，可提供單一讀數器，其能夠用於提供來自各種獨特組之不同檢定之結果。在此方面，每個盒可針對可執行之檢定之數目及類型來專門調適。舉例而言，對於特定檢定，可需要不同體積之樣本並且此可經由每個通道及/或腔室之適當設定尺寸來獨立地解決。因此，藉由增加或減少任何特定通道之尺寸，可增加或減少引入每個特定通道中之樣本之體積。此外，必要時，連接至一或多個通道之任何腔室之尺寸可增加或減少，視檢定之類型、步驟之數目及/或引入該通道中之樣本之體積而定。此容易藉由審閱本說明書之熟習此項技術者來判定。 The invention is further based on the development of an assay system comprising a disposable microfluidic cartridge and associated reader capable of performing a plurality of different assays for a single sample, the reader being capable of detecting and/or determining a plurality of samples from a single sample The level of the analyte and the output to the user. The present invention also allows for the receipt of various disposable cassettes by a reader, each of which is capable of performing a unique set of different assays. In this way, a single reader can be provided that can be used to provide results from different assays of various unique groups. In this regard, each box can be specifically adapted to the number and type of executable checks. For example, for a particular assay, samples of different volumes may be required and this may be independently resolved via the appropriate set size of each channel and/or chamber. Thus, by increasing or decreasing the size of any particular channel, the volume of the sample introduced into each particular channel can be increased or decreased. In addition, the size of any chamber connected to one or more channels may be increased or decreased as necessary, depending on the type of assay, the number of steps, and/or the volume of the sample introduced into the channel. This is easily determined by those skilled in the art who have reviewed this specification.

因此，在另一態樣中，提供用於進行複數個不同檢定之自給式拋棄式微流控系統，該微流控盒包含：用於將液體樣本引入微流控盒中之樣本輸入埠；多個微流控通道；該等微流控通道中之每一者適於接收液體樣本之一部分並且能夠使用一或多個試劑來對於樣本之該部分進行一或多 個檢定，該等試劑在引入液體樣本之前存在於該等微流控通道中之每一者內；及其中每個微流控通道內之流體運動藉由微流控系統之兩個或兩個以上氣體填充腔室之壓縮及/或減壓來獨立地可控制，該等腔室各自與該等微流控通道中之一或多者流體連通。 Therefore, in another aspect, a self-contained disposable microfluidic system for performing a plurality of different assays is provided, the microfluidic cartridge comprising: a sample input port for introducing a liquid sample into the microfluidic cartridge; a microfluidic channel; each of the microfluidic channels being adapted to receive a portion of a liquid sample and capable of performing one or more assays on the portion of the sample using one or more reagents, the reagents being introduced a liquid sample is previously present in each of the microfluidic channels; and fluid movement within each of the microfluidic channels is compressed by a chamber filled by two or more gases of the microfluidic system / or reduced pressure to be independently controllable, each of the chambers being in fluid communication with one or more of the microfluidic channels.

應瞭解本發明之上述進一步態樣可附加於或代替先前如上所述之態樣及實施方式。因此，關於本發明之先前態樣所描述之全部特徵可同樣適用於直接上述態樣並且由此可作為限制性或任擇特徵來包括在內。 It will be appreciated that the above-described further aspects of the invention may be added to or substituted for the aspects and embodiments previously described above. Accordingly, all of the features described in relation to the prior aspects of the invention may be equally applicable to the above-described aspects and thus may be included as a limitation or optional feature.

亦可使用本發明之盒來執行檢定，該等盒除了具有與一或多個腔室或腔室流體連通之通道以外，進一步包含不與一或多個氣體腔室中之任一者流體連通的一或多個通道。此檢定之非限制性實例在本文以下實例部分中描述。 The assay may also be performed using the cartridge of the present invention, which in addition to having a passage in fluid communication with one or more chambers or chambers, further comprising not in fluid communication with any of the one or more gas chambers One or more channels. Non-limiting examples of such assays are described in the Examples section below.

本發明之微流控盒就以下而言係自給式的：在執行每個檢定過程之前，除了樣本本身以外，進行每個檢定所需要的所有其他實體試劑存在於微流控盒中。因此，其他試劑，諸如反應物質、緩衝液、清洗液體等在檢定過程期間不引入盒中。通常，進入盒之唯一液體係液體樣本本身。可置放在該/每個通道中之任何試劑可最初經由液體來施加，但是其已經乾燥並且在進行任何特定檢定之前，盒可被視為乾燥的，並且沒有或實質上沒有液體存在。 The microfluidic cartridge of the present invention is self-contained in that all other physical reagents required for each assay are present in the microfluidic cartridge, except for the sample itself, prior to performing each assay procedure. Therefore, other reagents such as a reaction substance, a buffer, a washing liquid, and the like are not introduced into the cartridge during the assay process. Typically, the only liquid system liquid sample that enters the cartridge itself. Any reagent that can be placed in the/each channel can be initially applied via a liquid, but it has dried and the cartridge can be considered dry before and for any particular assay, and no or substantially no liquid is present.

如下論述，可向來自相關聯讀數器之盒提供加熱/冷卻及/或磁力施加，但是此不被理解為實體試劑。 As discussed below, heating/cooling and/or magnetic application may be provided to the cartridge from the associated reader, but this is not to be understood as a physical reagent.

在本發明之上下文中，多重檢定理解為意味著每個微流控盒不僅能夠執行來自引入盒中之單一樣本之複數個檢定，而且盒能夠執行複數個明顯不同類型之檢定。舉例而言，本發明之每個微流控盒能夠執行以下類型之檢定中之至少兩者、三者、四者、五者或更多者：免疫檢定、核酸檢定、基於受體的檢定、競爭檢定、細胞計數檢定、比色檢定、酶檢定、電泳檢定、電化學檢定、光譜檢定、層析檢定、顯微鏡檢定、局部檢定、量熱檢定、濁度檢定、凝集檢定、黏度檢定、凝血檢定、凝結時間檢定、蛋白質合成檢定、組織學檢定、培養檢定、滲透壓、化學、生物化學、離子、氣體或吸收檢定。在某些實施方式中，可執行特定類型之檢定以便偵測不同分析物。舉例而言，可執行一個以上免疫檢定以便偵測不同分析物。該一個以上免疫檢定可在單一及/或多個微流控通道中執行。 In the context of the present invention, multiple verification is understood to mean that each microfluidic cartridge is capable of performing not only a plurality of assays from a single sample introduced into the cartridge, but also that the cartridge is capable of executing a plurality of distinctly different types of assays. For example, each microfluidic cartridge of the present invention is capable of performing at least two, three, four, five or more of the following types of assays: immunoassays, nucleic acid assays, receptor-based assays, Competition assay, cell count assay, colorimetric assay, enzyme assay, electrophoresis assay, electrochemical assay, spectral assay, chromatographic assay, microscopic assay, local assay, calorimetric assay, turbidity assay, agglutination assay, viscosity assay, coagulation assay , coagulation time assay, protein synthesis assay, histological assay, culture assay, osmotic pressure, chemistry, biochemistry, ion, gas or absorption assay. In certain embodiments, a particular type of assay can be performed to detect different analytes. For example, more than one immunoassay can be performed to detect different analytes. The one or more immunoassays can be performed in a single and/or multiple microfluidic channels.

在一實施方式中，本發明之微流控盒被設計來進行與特定疾病或病狀有關的一組檢定。示例性測試組可包括針對以下各項之多組檢定：心臟病狀、腎上腺病狀、肝功能、腎功能、神經功能、糖尿病、妊娠及妊娠病狀、代謝病狀及藥物濫用。 In one embodiment, the microfluidic cartridge of the present invention is designed to perform a set of assays associated with a particular disease or condition. Exemplary test groups can include multiple sets of assays for: heart disease, adrenal gland conditions, liver function, renal function, neurological function, diabetes, pregnancy and pregnancy conditions, metabolic conditions, and drug abuse.

舉例而言，被設計來檢定與心臟病狀相關聯之標誌物的微流控盒可包含被設計來偵測及/或判定以下中之一或多者之水準的一或多個檢定：脂質概況，其可偵測例如低密度脂蛋白(LDL)、高密度脂蛋白(HDL)、甘油三酯及/或總膽固醇；脂蛋白元，亦即脂蛋白之蛋白質組分，不包括在標準脂質概況中，但是可單獨地測試。異常水準可促進動脈粥樣硬化，並且可增加冠 狀動脈疾病(CAD)及中風之風險；升半胱胺酸係一種胺基酸(蛋白質結構單元)。較高血液水準可促進動脈粥樣硬化及CAD，以及可導致心臟病發作或中風之血液凝塊；肌鈣蛋白；BNP；C-反應蛋白(CRP)係反映低水準之全身炎症並且在處在CAD之風險中之人中增加的物質；及心臟標誌物，諸如心臟酶研究量測某些酶，諸如CK-MB或肌鈣蛋白，或心臟激素諸如大腦利尿鈉肽，該等心臟激素在心臟受脅迫或患病或由於心臟病發作而受損時釋放。 For example, a microfluidic cartridge designed to characterize a marker associated with a heart condition can include one or more assays designed to detect and/or determine the level of one or more of the following: lipid Overview, which can detect, for example, low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglyceride, and/or total cholesterol; lipoprotein, a protein component of lipoproteins, not included in standard lipids In the profile, but can be tested separately. Abnormal levels promote atherosclerosis and increase the risk of coronary artery disease (CAD) and stroke; cysteine is an amino acid (protein building block). Higher blood levels can promote atherosclerosis and CAD, as well as blood clots that can lead to heart attacks or strokes; troponin; BNP; C-reactive protein (CRP) reflects low-level systemic inflammation and is at Increased substances in people at the risk of CAD; and cardiac markers such as cardiac enzymes to measure certain enzymes, such as CK-MB or troponin, or cardiac hormones such as brain natriuretic peptides, in the heart Released when stressed or sick or damaged by a heart attack.

經歷應力或其他病狀之受試者可經受腎上腺功能的一組檢定，該組可包括以下各項中之一或多者：醛固酮控制身體中之鹽、鉀及水分平衡並且有助於調節血壓。此激素之過度產生(醛固酮過多症)或產生不足(醛固酮減少症)可藉由腫瘤或腎上腺內之其他異常(原發性；例如，腎上腺癌症)導致或可由腎上腺以外之問題(繼發性)導致；皮質醇係一種糖皮質激素，其有助於控制碳水化合物、蛋白質及脂肪之代謝；介導身體對於應力之響應；並且調節免疫系統。最經常藉由良性腎上腺瘤造成的皮質醇之過度分泌導致庫欣氏症候群。分泌不足可指示被稱為艾迪生氏病的一種腎上腺機能不全形式。通常量測血液水準及尿液水準(被稱為游離皮質醇)；18-羥皮質醇，一種皮質醇代謝之產物，係在患有原發性醛固酮過多症之患者中過量產生的異常類固醇。量測此激素之血液水準可有 助於判定是否原發性醛固酮過多症藉由被稱為腎上腺瘤之腫瘤，或腎上腺組織之過度生長(增生)導致；在患有腺瘤之人中之水準顯著較高；及DHEA-S，或脫氫表雄酮硫酸鹽，一種藉由腎上腺合成之性激素(雄性激素)，係睾酮之前驅物。在女性中，腎上腺係雄性激素之主要及有時唯一來源。較高DHEA-S水準與男性化(男性身體特性)、多毛症(過度毛髮生長)、無月經(缺乏月經)及***相關聯。腎上腺異常諸如腫瘤可導致異常高DHEA-S水準。 A subject undergoing stress or other condition may undergo a set of tests for adrenal function, which may include one or more of the following: aldosterone controls salt, potassium and water balance in the body and helps regulate blood pressure . Excessive production of this hormone (hyperketidemia) or underproduction (aldosteronism) can be caused by tumors or other abnormalities in the adrenal gland (primary; for example, adrenal cancer) or by problems other than the adrenal gland (secondary) Cortisol is a glucocorticoid that helps control the metabolism of carbohydrates, proteins, and fats; mediates the body's response to stress; and regulates the immune system. Cushing's syndrome is most often caused by excessive secretion of cortisol by benign adrenal adenomas. Insufficient secretion may indicate an incomplete form of adrenal insufficiency known as Addison's disease. Blood levels and urine levels (known as free cortisol) are usually measured; 18-hydroxycortisol, a product of cortisol metabolism, is an overdose of abnormal steroids in patients with primary aldosteronism. Measuring the blood level of this hormone can help determine whether primary aldosteronism is caused by a tumor called adrenal adenoma or excessive growth (proliferation) of adrenal tissue; the level of a person with adenoma Significantly higher; and DHEA-S, or dehydroepiandrosterone sulfate, a sex hormone synthesized by the adrenal gland (androgen), a testosterone precursor. In women, the main and sometimes the only source of adrenal androgen. Higher DHEA-S levels are associated with masculinity (male body characteristics), hirsutism (excessive hair growth), menstruation (lack of menstruation), and infertility. Adrenal abnormalities such as tumors can result in abnormally high DHEA-S levels.

肝功能測試用於幫助判定可歸因於肝臟疾病之諸如黃疸之症狀之原因。其亦用於在例如酗酒者或暴露於肝炎病毒之人中篩檢可能肝損傷，並且亦監測異常肝臟功能之變化。因此，本發明之肝臟功能微流控盒可包括以下各項中之一或多者：酶測試：肝臟係藉由許多酶來控制之許多生化反應之部位，該等酶包括丙胺酸轉胺酶(ALT)、天門冬胺酸轉胺酶(AST)、鹼性磷酸酶(ALP)及γ麩胺醯基轉移酶(GGT)。血流中之肝臟酶之較高水準可指示肝損傷；然而，其不一定指向特定肝臟疾病。雖然酶測試可個別地安排，但是其在組合執行時提供更多資訊，因為在影響其他器官之疾病中，許多肝臟酶之水準可升高；膽紅素，膽汁中之主要色素，係血紅蛋白之分解產物，該血紅蛋白係紅血球中之含鐵物質。通常，僅少量膽紅素在血液中循環。較高血液水準可由許多形式之肝臟及膽道疾病，包括肝炎及膽管阻塞所導致。血液中之過量膽紅素之存在產生皮膚及眼睛之微黃色變色，被稱為黃疸；白蛋白係如同血流中之大多數蛋白質一樣，藉由肝臟來合成 的主要蛋白質。血清(在全血凝結之後保留的血液之液體部分)中之白蛋白之水準減少係慢性肝臟疾病之指示；凝血酶原時間(PT)係可執行來評價肝臟之功能的血液凝結研究。因為凝血酶原係藉由肝臟合成之凝血蛋白質中之一者，所以異常PT可反映肝臟功能障礙；病毒性肝炎測試可在具有異常肝臟酶之人中進行，其病史及/或症狀疑似患有疾病。(症狀包括低燒、不適、食欲損失及疲勞，但是不一定存在。)在美國發現的三種最常見類型之此病毒係A型、B型及C型肝炎(被稱為HAV、HBV及HCV)；其全部藉由測試僅在受感染個體之血液中發現之特定抗原或抗體之存在來偵測。不同抗體/抗原測試可執行，視肝炎類型疑似。另外，特定抗體之存在可表示是否感染處於急性或慢性階段。 Liver function tests are used to help determine the cause of symptoms such as jaundice attributable to liver disease. It is also used to screen for possible liver damage in, for example, alcoholics or people exposed to hepatitis virus, and also to monitor changes in abnormal liver function. Thus, the liver function microfluidic cassette of the present invention may comprise one or more of the following: enzyme testing: the liver is a site of many biochemical reactions controlled by a number of enzymes, including alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and gamma glutamine thiotransferase (GGT). A higher level of liver enzymes in the bloodstream may indicate liver damage; however, it does not necessarily point to a particular liver disease. Although the enzyme test can be arranged individually, it provides more information when the combination is performed, because many liver enzymes can be elevated in diseases affecting other organs; bilirubin, the main pigment in bile, is hemoglobin Decomposition product, the hemoglobin is an iron-containing substance in red blood cells. Usually, only a small amount of bilirubin circulates in the blood. Higher blood levels can result from many forms of liver and biliary tract disease, including hepatitis and biliary obstruction. The presence of excess bilirubin in the blood produces a yellowish discoloration of the skin and eyes, known as jaundice; albumin is the main protein synthesized by the liver, like most proteins in the bloodstream. The reduction in albumin levels in serum (the liquid portion of the blood retained after whole blood clotting) is indicative of chronic liver disease; prothrombin time (PT) is a blood coagulation study that can be performed to assess liver function. Because prothrombin is one of the coagulation proteins synthesized by the liver, abnormal PT can reflect liver dysfunction; viral hepatitis test can be performed in people with abnormal liver enzymes, whose history and/or symptoms are suspected to have disease. (Symptoms include low fever, discomfort, loss of appetite, and fatigue, but not necessarily.) The three most common types of viruses found in the United States are Type A, Type B, and Type C (known as HAV, HBV, and HCV); All of them are detected by testing for the presence of specific antigens or antibodies found only in the blood of infected individuals. Different antibody/antigen tests can be performed, depending on the type of hepatitis suspected. Additionally, the presence of a particular antibody can indicate whether the infection is in an acute or chronic phase.

多組測試經常用於考慮受試者患上糖尿病之風險或確認受試者患有I或II型糖尿病。與如上所述之脂質組一樣，糖尿病組微流控盒可被設計來進行以下檢定中之一或多者：血液病症諸如感染或貧血之全血計數(CBC)測試；禁食葡萄糖用於偵測高血糖及低血糖，以幫助診斷糖尿病，並且監測患有糖尿病之人的葡萄糖水準；血紅蛋白A1c可偵測糖尿病前期，對它進行診斷，或發現是否糖尿病處在控制下；並且糖尿病尿分析判定若在你的尿液中發現白蛋白(蛋白質)(若發現，則可能受試者之腎臟不能正常地工作)。 Multiple sets of tests are often used to consider the risk of diabetes in a subject or to confirm that a subject has type I or type 2 diabetes. Like the lipid group described above, the diabetic group microfluidic cartridge can be designed to perform one or more of the following assays: blood disorders such as infection or anemia of whole blood count (CBC) test; fasting glucose for detection High blood sugar and hypoglycemia to help diagnose diabetes and monitor the glucose level of people with diabetes; hemoglobin A1c can detect pre-diabetes, diagnose it, or find out if diabetes is under control; and diabetes urinalysis If albumin (protein) is found in your urine (if found, the subject's kidneys may not work properly).

亦可測試藥物濫用或被視為運動員及女性禁用的藥物。被設 計來偵測及/或判定受試者中之藥物濫用水準的本發明之微流控盒可被設計來檢定以下中之一或多者：***；巴比妥酸鹽；丁基原啡因；苯二氮平；古柯鹼；搖頭丸；甲基***；***(鴉片劑/嗎啡)；***；三環抗抑鬱藥；***及/或其他精神作用劑 Drug abuse or drugs that are considered to be banned by athletes and women can also be tested. The microfluidic cartridge of the present invention designed to detect and/or determine the level of drug abuse in a subject can be designed to assay one or more of the following: amphetamine; barbiturate; butylmorphine; Benzodiazepine; ***e; ecstasy; methamphetamine; heroin (opiate/morphine); methadone; tricyclic antidepressant; cannabis and/or other psychoactive agents

應認識到以上描述多組檢定僅僅係示範性並且不應被視為具有限制性。根據本發明，可設想多組特定檢定並且提供根據本發明之盒以便進行該組特定檢定。 It will be appreciated that the various sets of assays described above are merely exemplary and should not be considered as limiting. In accordance with the present invention, multiple sets of specific assays are contemplated and a cartridge in accordance with the present invention is provided for performing the set of specific assays.

雖然每一組檢定在本發明之微流控盒內進行，但是每個檢定之結果需要偵測及/或判定。此藉由如本文描述之讀數器來執行。 While each set of assays is performed within the microfluidic cartridge of the present invention, the results of each assay require detection and/or determination. This is performed by a reader as described herein.

讀數器可包括盒判定構件，其可為存在於讀數器中之條形碼/QR代碼讀數器等，其被設計來讀取條形碼/QR代碼或微流控盒上之其他類型之代碼。代碼向讀數器傳達關於微流控盒之類型及將要進行之檢定的資訊，以使得讀數器準備執行及偵測/判定來自特定微流控盒之結果。在更簡單實施方式中，讀數器之該等接收埠可被設計來接受僅特定微流控盒類型，很像鎖及鑰匙。因此，每個接收埠可僅接收特定類型之盒，其中將盒引入特定接收埠中向讀數器提供關於所***之盒之類型及將要進行之檢定的指示。使用者亦可將詳細資料輸入讀數器中以使得向讀數器提供關於將要執行之檢定的指示，但是此可能不太理想的，因為其可導致使用者錯誤。 The reader may include a cartridge determination component, which may be a barcode/QR code reader or the like present in the reader, designed to read the barcode/QR code or other type of code on the microfluidic cartridge. The code communicates information to the reader regarding the type of microfluidic cartridge and the assay to be performed to prepare the reader for execution and to detect/determine results from a particular microfluidic cartridge. In a simpler embodiment, the receivers of the reader can be designed to accept only certain microfluidic cartridge types, much like locks and keys. Thus, each receiving cassette can only receive a particular type of cartridge, wherein the introduction of the cartridge into a particular receiving cassette provides an indication to the reader regarding the type of cartridge inserted and the assay to be performed. The user may also enter the details into the reader to provide an indication to the reader regarding the check to be performed, but this may not be ideal as it may result in user error.

本發明之讀數器被構建成使得其能夠接收複數個不同微流控盒。「不同」應理解為意味著本發明之盒可適於進行一組特定檢定，而不是盒在視覺上表觀明顯不同。亦即，兩個盒在並排放置時可在視覺上看起 來非常相似的，但是舉例而言，一個盒可適於執行適合於偵測心臟病的一組檢定並且另一個盒可適於執行適合於糖尿病偵測的一組檢定。 The reader of the present invention is constructed such that it can receive a plurality of different microfluidic cartridges. "Different" is understood to mean that the cartridge of the present invention can be adapted to perform a particular set of assays, rather than the box being visually apparently distinct. That is, the two boxes may look very similar when placed side by side, but for example, one box may be adapted to perform a set of tests suitable for detecting heart disease and another box may be adapted to perform the appropriate A set of tests for diabetes detection.

因此在另一態樣中，提供用於進行多組檢定的多重檢定平臺，該多重檢定平臺包含複數個微流控盒，每個盒能夠對於樣本進行一組規定檢定及讀數器，該讀數器被構建來能夠接收及檢驗該等複數個微流控盒中之每一者，其中讀數器可組配來偵測及/或判定可存在於樣本中之一組分析物的水準。 Thus, in another aspect, a multi-assay platform for performing a plurality of sets of assays is provided, the multi-assay platform comprising a plurality of microfluidic cartridges, each cartridge being capable of performing a set of prescribed assays and readers for the sample, the reader It is constructed to receive and test each of the plurality of microfluidic cartridges, wherein the readers can be configured to detect and/or determine the level of analytes that can be present in the sample.

在使用中，受試者預定使用一組特定檢定來測試，或患者訪問健康照護提供者，諸如醫生、護士或其他醫療專業人員並且健康照護提供者將受試者確認為需要進行一組適當測試。患者或健康照護提供者選擇盒，其被組配來執行該組預期檢定並且將此選定盒***讀數器中。讀數器從存在於盒上之特徵來判定盒被設計來進行哪一組檢定並且讀數器將自身適當地組配以便能夠運行檢定並偵測及/或判定存在於來自受試者之樣本中之一組特定分析物之水準。提供或獲得來自受試者之樣本並且將樣本引入盒之輸入埠中。藉由讀數器及盒在一起運作，對於樣本進行該組檢定，並且在檢定結束時，讀數器偵測及/或判定存在於樣本中之分析物之水準。然後，讀數器將該組檢定之結果提供給受試者及/或健康照護提供者。 In use, the subject is scheduled to be tested using a specific set of tests, or the patient visits a health care provider, such as a doctor, nurse or other medical professional and the health care provider confirms the subject as needing a suitable set of tests . The patient or health care provider selects a cartridge that is assembled to perform the set of expected assays and inserts the selected cartridge into the reader. The reader determines from the features present on the cartridge which set of assays the cartridge is designed to perform and the readers properly assemble themselves to be able to run the assay and detect and/or determine that it is present in the sample from the subject. The level of a specific set of analytes. A sample from the subject is provided or obtained and the sample is introduced into the input port of the cassette. The set of assays is performed on the sample by the reader and the cartridge operating together, and at the end of the assay, the reader detects and/or determines the level of analyte present in the sample. The reader then provides the results of the set of assays to the subject and/or health care provider.

除了健康照護提供者以外，使用者可為執法官員，或運動藥物測試官員，例如，當受試者係針對例如不適當的藥物使用來進行測試的個體時。 In addition to the health care provider, the user may be a law enforcement officer, or a sports drug testing officer, for example, when the subject is an individual who is tested for, for example, inappropriate drug use.

本發明現在參考以下編號條款來進一步限定： The invention is now further defined by reference to the following numbering clauses:

1.一種用於對於一液體樣本進行一檢定的微流控盒，該微流控盒包含 連接至至少一個微流控通道的一樣本輸入埠，其中每個/該微流控通道包含置放在其中的用於進行該檢定之一或多個試劑及一偵測區，每個/該微流控通道進一步流體連接至一可壓縮氣體填充腔室，其中將該腔室之一外表面壓縮或減壓相應地導致氣體從該腔室中排出或抽吸至該腔室中，進而導致該/每個微流控通道內之該液體樣本之一往復式運動。 CLAIMS 1. A microfluidic cartridge for performing an assay on a liquid sample, the microfluidic cartridge comprising an identical input port coupled to at least one microfluidic channel, wherein each/the microfluidic channel comprises a placement In each of the one or more reagents and a detection zone for performing the assay, each/the microfluidic channel is further fluidly coupled to a compressible gas-filled chamber, wherein an outer surface of one of the chambers is compressed Or decompression correspondingly causes gas to be expelled or drawn into the chamber from the chamber, thereby causing reciprocating movement of one of the liquid samples within the/microfluidic channel.

2.如條款1之微流控盒，其中在該液體樣本與置放在該/每個微流控通道內之該一或多個試劑反應之後，從該腔室排出之氣體用來將液體從該/每個微流控通道內之該偵測區中移除，以使得可在一實質上無液體環境中偵測該/每個偵測區內之任何分析物或分析物反應產物。 2. The microfluidic cartridge of clause 1, wherein the gas discharged from the chamber is used to liquid after the liquid sample is reacted with the one or more reagents placed in the/each microfluidic channel The detection zone is removed from the/each microfluidic channel such that any analyte or analyte reaction product within the/detection zone can be detected in a substantially liquid-free environment.

3.如條款1或2之微流控盒，其包含複數個微流控通道，其中該等複數個微流控通道中之每一者與該樣本輸入埠流體連通。 3. The microfluidic cartridge of clause 1 or 2, comprising a plurality of microfluidic channels, wherein each of the plurality of microfluidic channels is in fluid communication with the sample input port.

4.如條款3之微流控盒，其中該等複數個微流控通道中之每一者連接至一相應氣體填充腔室，且/或兩個或兩個以上微流控通道連接至一氣體填充腔室。 4. The microfluidic cartridge of clause 3, wherein each of the plurality of microfluidic channels is connected to a respective gas filled chamber, and/or two or more microfluidic channels are connected to one The gas fills the chamber.

5.如前述條款中任一項之微流控盒，其中該樣本埠連接至該/每個微流控通道之一第一端並且該/每個微流控通道之一第二端連接至該等氣體填充腔室中之一或多者。 5. The microfluidic cartridge of any of the preceding clauses, wherein the sample cartridge is coupled to one of the first ends of the/each microfluidic channel and the second end of the/each microfluidic channel is coupled to The gases fill one or more of the chambers.

6.如前述條款中任一項之微流控盒，其進一步包含被設計來接收流體廢物及/或過量液體樣本的一或多個吸收器特徵。 6. The microfluidic cartridge of any of the preceding clauses, further comprising one or more absorber features designed to receive fluid waste and/or excess liquid sample.

7.如前述條款中任一項之微流控盒，其中該盒及安置在其中之該等通道及其他特徵藉由三個分離平面基板之一層疊物來形成，該層疊物包含一頂部基板、一底部基板及安置在該頂部與底部基板之間的中間基板。 7. The microfluidic cartridge of any of the preceding clauses, wherein the cartridge and the channels and other features disposed therein are formed by a laminate of three separate planar substrates, the laminate comprising a top substrate a bottom substrate and an intermediate substrate disposed between the top and bottom substrates.

8.如條款7之微流控盒，其中每個層具有一均勻厚度並且由相同材料形成，視需要每個層具有相同均勻厚度。 8. The microfluidic cartridge of clause 7, wherein each layer has a uniform thickness and is formed from the same material, each layer having the same uniform thickness as desired.

9.如條款7或8中任一項之微流控盒，其中該盒由一捲筒或卷對卷過程來形成。 9. The microfluidic cartridge of any of clauses 7 or 8, wherein the cartridge is formed by a roll or roll-to-roll process.

10.如條款7-9中任一項之微流控盒，其中該等平面基板藉由施加熱及/或使用黏著劑來密封在一起。 10. The microfluidic cartridge of any of clauses 7-9, wherein the planar substrates are sealed together by applying heat and/or using an adhesive.

11.如條款10之微流控盒，其中該等平面基板使用一黏著劑來密封在一起，該黏著劑有彈性並且促進每個/該腔室之可壓縮性。 11. The microfluidic cartridge of clause 10, wherein the planar substrates are sealed together using an adhesive that is resilient and promotes compressibility of each/the chamber.

12.如前述條款中任一項之微流控盒，其中該盒中之該/每個微流控通道包含一或多個流體止擋特徵，其被設計來防止該樣本及/或其他流體僅藉助於毛細管作用來經過該止擋特徵。 The microfluidic cartridge of any of the preceding clauses, wherein the/each microfluidic channel in the cartridge comprises one or more fluid stop features designed to prevent the sample and/or other fluids This stop feature is only passed by means of capillary action.

13.如前述條款中任一項之微流控盒，其包含一單向閥，該閥被設計來僅在藉由毛細管作用將一液體樣本引入該盒中時允許氣體退出該盒，同時不允許經由該閥將流體引入該盒中。 13. The microfluidic cartridge of any of the preceding clauses, comprising a one-way valve designed to allow gas to exit the cartridge only when a liquid sample is introduced into the cartridge by capillary action, while not Fluid is allowed to be introduced into the cartridge via the valve.

14.如條款13之微流控盒，其中該閥相鄰於一止擋特徵來定位，該止擋特徵被設計來防止該樣本僅藉由毛細管作用在該微流控通道內進一步傳送。 14. The microfluidic cartridge of clause 13, wherein the valve is positioned adjacent to a stop feature that is designed to prevent further transfer of the sample within the microfluidic channel by capillary action only.

15.如條款14之微流控盒，其中該閥定位在具有比該/每個微流控通道更小之尺寸的一微流控通道中並且與該等微流控通道中之一者流體連通。 15. The microfluidic cartridge of clause 14, wherein the valve is positioned in a microfluidic channel having a smaller size than the/each microfluidic channel and is fluid with one of the microfluidic channels Connected.

16.如前述條款中任一項之微流控盒，其包含與該/每個通道接觸之一或多個電極特徵，其用於量測或偵測存在於該/每個通道中之液體。 16. The microfluidic cartridge of any of the preceding clauses, comprising one or more electrode features in contact with the/each channel for measuring or detecting liquid present in the/each channel .

17.如前述條款中任一項之微流控盒，其進一步包含置放在該通道內之 一分析物結合劑，其中視需要該分析物結合劑結合至該通道之一表面。 17. The microfluidic cartridge of any of the preceding clauses, further comprising an analyte binding agent disposed within the channel, wherein the analyte binding agent is bound to a surface of the channel as desired.

18.如條款17之微流控盒，其中該結合劑附接至一磁性或順磁粒子。 18. The microfluidic cartridge of clause 17, wherein the binding agent is attached to a magnetic or paramagnetic particle.

19.如條款17或18之微流控盒，其中該結合劑或磁性/順磁粒子置放在該盒之該/每個微流控通道內，以使得在該樣本施加至該盒並且被抽吸至該/每個通道中之後，該等黏合劑或磁性/順磁粒子藉由該液體樣本來再懸浮。 19. The microfluidic cartridge of clause 17 or 18, wherein the binder or magnetic/paramagnetic particles are placed in the/each microfluidic channel of the cartridge such that the sample is applied to the cartridge and is After being drawn into the/each channel, the binder or magnetic/paramagnetic particles are resuspended by the liquid sample.

20.如條款17-19中任一項之微流控盒，其中該結合劑或磁性/順磁粒子置放在該/每個微流控通道之一區域內，該區域藉由在該置放區域之任一端處之特徵來界定，該等特徵被設計來在該等磁性/順磁粒子最初置放在該/每個通道中時限制該等粒子之運動。 The microfluidic cartridge of any one of clauses 17 to 19, wherein the binder or magnetic/paramagnetic particles are placed in an area of the/each microfluidic channel, the region being The features at either end of the discharge zone are defined, and the features are designed to limit the movement of the magnetic/paramagnetic particles as they are initially placed in the/each channel.

21.如條款19或20中任一項之微流控盒，其中該等磁性/順磁粒子置放在與一磁鐵或磁力緊密鄰近之該盒之表面相反的該/每個通道之一表面上。 The microfluidic cartridge of any one of clauses 19 or 20, wherein the magnetic/paramagnetic particles are placed on a surface of the/each channel opposite the surface of the cartridge in close proximity to a magnet or magnetic force on.

22.如前述條款中任一項之微流控盒，其中該盒進一步包含置放在該/每個微流控通道內之一或多個額外試劑，該等額外試劑促進存在於該樣本中之分析物之偵測。 The microfluidic cartridge of any of the preceding clauses, wherein the cartridge further comprises one or more additional reagents disposed in the/each microfluidic channel, the additional reagents being facilitated to be present in the sample Detection of analytes.

23.如條款22之微流控盒，其中該一或多個額外試劑包括一標記物，該標記物適於特異性結合至將要偵測之一分析物以促進分析物偵測。 23. The microfluidic cartridge of clause 22, wherein the one or more additional reagents comprise a label adapted to specifically bind to one of the analytes to be detected to facilitate analyte detection.

24.如條款22或23之微流控盒，其中經由將氣體抽吸回到該/每個氣體填充腔室中，分析物在該/每個微流控通道之一第一區域中結合至該分析物結合劑，然後被傳送至該/每個微流控通道之另外一或多個區域，其中置放該一或多個其他試劑及/或標記物。 24. The microfluidic cartridge of clause 22 or 23, wherein the analyte is bound to the first region of the one/each microfluidic channel by pumping gas back into the/each gas-filled chamber The analyte binding agent is then delivered to one or more additional regions of the per microfluidic channel in which the one or more additional reagents and/or labels are placed.

25.如前述條款中任一項之微流控盒，其中該盒能夠對於一單一樣本執行複數個(諸如2、3、4、5、6、7、8、9、10或更多個)相同及/或不同檢定。 The microfluidic cartridge of any of the preceding clauses, wherein the cartridge is capable of performing a plurality (such as 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) for a single sample Same and / or different tests.

26.如前述條款中任一項之微流控盒，其中施加至該盒之該樣本之體積小於50μl，諸如小於40μl、30μl或20μl。 The microfluidic cartridge of any of the preceding clauses, wherein the sample applied to the cartridge has a volume of less than 50 μl , such as less than 40 μl , 30 μl or 20 μl .

27.一種套組，其包含如前述條款中任一項之微流控盒，以及一樣本收集裝置。 27. A kit comprising a microfluidic cartridge according to any of the preceding clauses, and the same collection device.

28.如條款26之套組，其中該樣本收集裝置適於***該盒之樣本輸入埠並且其後為該輸入埠提供一密封。 28. The kit of clause 26, wherein the sample collection device is adapted to insert a sample input port of the cartridge and thereafter provide a seal for the input port.

29.如條款27之套組，其用於進行一核酸偵測檢定。 29. The kit of clause 27 for performing a nucleic acid detection assay.

30.一種與如條款1-26中任一項之微流控盒，或如條款26-29之套組一起使用的讀數器裝置，該讀數器裝置包含：用於將該微流控盒引入該讀數器裝置中之一接收埠；力施加構件，該力施加構件用於接觸該盒之該/每個氣體填充腔室之一外表面並且能夠將一可變力施加至該/每個氣體填充腔室，其中最初將力施加至該/每個氣體填充腔室之表面導致氣體從該/每個氣體填充腔室並且沿著該/每個微流控通道排出遠離該/每個腔室；並且施加至該/每個氣體填充腔室之該力之減少導致該/每個微流控通道內之氣體被抽吸回到並且進入該氣體填充腔室；及偵測構件，該偵測構件用於實現存在於引入該微流控盒中之一液體樣本內之一所需分析物或分析物反應產物的偵測。 30. A reader device for use with a microfluidic cartridge according to any of clauses 1-26, or a kit according to clauses 26-29, the reader device comprising: for introducing the microfluidic cartridge One of the reader devices receives a weir; a force applying member for contacting an outer surface of the/each gas-filled chamber of the cartridge and capable of applying a variable force to the/each gas Filling the chamber, wherein initially applying a force to the surface of the/each gas-filled chamber causes gas to be evacuated from the/each gas-filled chamber and along the/each microfluidic channel away from the/each chamber And the reduction in the force applied to the/each gas-filled chamber causes the gas in the/each microfluidic channel to be drawn back into and into the gas-filled chamber; and the detection member, the detection The means is for effecting detection of an analyte or analyte reaction product present in one of the liquid samples introduced into the microfluidic cartridge.

31.如條款30之讀數器裝置，其包含適於接收不同大小之盒的一接收埠，每個不同大小之盒被設計來執行規定數目之檢定。 31. The reader device of clause 30, comprising a receiving cassette adapted to receive cartridges of different sizes, each of the different sized cartridges being designed to perform a prescribed number of assays.

32.如條款31之讀數器裝置，其中該接收埠經如此調適以確保每個不同大小之盒的正確***及識別。 32. The reader device of clause 31, wherein the receiving cassette is adapted to ensure proper insertion and identification of each of the different sized cartridges.

33.如條款30-32之讀數器裝置，其進一步包含緊密鄰近被引入該讀數器中之如條款18-26之盒的一永磁鐵(或被設計來向該盒施加一磁場的電磁鐵)，以便該等磁性/順磁粒子集中並保持在該盒之該/每個微流控通道之偵測區中。 33. The reader device of clauses 30-32, further comprising a permanent magnet (or an electromagnet designed to apply a magnetic field to the cartridge) in close proximity to the cartridge of clauses 18-26 introduced into the reader, The magnetic/paramagnetic particles are concentrated and held in the detection zone of the/each microfluidic channel of the cartridge.

34.如條款30-33之讀數器裝置，其中該力施加構件呈一指狀物或一底座形式，其被設計來接觸該盒之一腔室之一外表面並且向其施加力。 34. The reader device of clause 30-33, wherein the force applying member is in the form of a finger or a base that is designed to contact and apply a force to an outer surface of one of the chambers of the cartridge.

35.如條款34之讀數器裝置，其中該指狀物/底座被設計來僅接觸氣體填充腔室之該總外表面之一部分。 35. The reader device of clause 34, wherein the finger/base is designed to contact only a portion of the total outer surface of the gas-filled chamber.

36.如條款35之讀數器裝置，其中每個指狀物/底座經設定大小來接觸每個腔室之10與50%之間之外表面。 36. The reader device of clause 35, wherein each finger/base is sized to contact between 10 and 50% of the surface of each chamber.

37.如條款30-36中任一項之讀數器裝置，其中該力施加構件被設計來使用存在於該讀數器內之一電動機來升高及降低以與該盒之表面接觸。 The reader device of any of clauses 30-36, wherein the force applying member is designed to be raised and lowered using a motor present in the reader to contact the surface of the cartridge.

38.如條款37之讀數器裝置，其中該電動機能夠以一可變速率來升高及降低該力施加構件以使得該盒內之氣體可以不同速率被抽吸至該/每個氣體填充腔室及/或從其中排出。 38. The reader device of clause 37, wherein the motor is capable of raising and lowering the force applying member at a variable rate such that gas within the cartridge can be drawn to the/each gas filled chamber at a different rate And / or discharged from it.

39.如條款30-38之讀數器裝置，其中該偵測構件係一光學偵測裝置，諸如一螢光計或分光光度計。 39. The reader device of clause 30-38, wherein the detecting member is an optical detecting device such as a fluorometer or a spectrophotometer.

40.如條款30-39中任一項之讀數器裝置，其進一步包含允許在一特定溫度，或複數個溫度下進行檢定之加熱及/或冷卻構件。 40. The reader device of any of clauses 30-39, further comprising a heating and/or cooling member that allows for verification at a particular temperature, or a plurality of temperatures.

41.一種對於一液體樣本進行一檢定之方法，該方法包含：a)將如條款1-26中任一項之微流控盒引入如條款30-40中任一項之讀數器裝置中； b)向該微流控盒之一/該氣體填充腔室施加一力，以便將氣體之一部分從該腔室中排出；c)將一液體樣本引入該微流控盒中並且允許毛細管作用將該樣本抽吸至該微流控通道中，或減少施加至該氣體填充腔室之該力，以使得空氣被抽吸至該腔室中，導致一液體樣本被抽吸至該微流控通道中；d)減少施加至該微流控盒之該腔室之力，以使得空氣被抽吸至該腔室中，導致該液體樣本被進一步抽吸至該微流控通道中以便允許與一分析物結合劑及視需要一或多個進一步試劑接觸；e)允許在該微流控通道之一偵測區中形成並捕獲任何分析物/分析物結合劑複合物或分析物反應產物/分析物結合劑複合物；f)視需要增加施加至該微流控盒之該氣體填充腔室之力，以使得氣體從該腔室排出，導致液體從捕獲該分析物/分析物結合劑複合物之該微流控通道之至少一部分中排出，以使得所捕獲的分析物/分析物結合劑複合物存在於一實質上無液體環境中；及g)視需要在該實質上無液體環境中偵測任何所捕獲分析物或分析物反應產物。 41. A method of performing a assay on a liquid sample, the method comprising: a) introducing a microfluidic cartridge according to any one of clauses 1-26 into a reader device according to any one of clauses 30-40; b) applying a force to one of the microfluidic cartridges/the gas-filled chamber to expel a portion of the gas from the chamber; c) introducing a liquid sample into the microfluidic cartridge and allowing capillary action to Pumping the sample into the microfluidic channel or reducing the force applied to the gas-filled chamber such that air is drawn into the chamber, causing a liquid sample to be drawn to the microfluidic channel Medium; d) reducing the force applied to the chamber of the microfluidic cartridge such that air is drawn into the chamber, causing the liquid sample to be further drawn into the microfluidic channel to allow Analyte binding agent and, if desired, one or more further reagent contacts; e) allowing formation and capture of any analyte/analyte binding agent complex or analyte reaction product/analysis in one of the microfluidic channel detection zones Binding agent complex; f) added to the microfluidic cartridge as needed The gas fills the force of the chamber such that gas is expelled from the chamber, causing liquid to drain from at least a portion of the microfluidic channel that captures the analyte/analyte binder complex, such that the captured analysis The analyte/analyte binder complex is present in a substantially liquid free environment; and g) detecting any captured analyte or analyte reaction product in the substantially liquid free environment as desired.

42.如條款41之方法，其中步驟d)作為單一或多個步驟來執行，其中對應於力之減少執行次數，該樣本相應地被抽吸至該/每個微流控通道內之另一個或多個連續位置。 42. The method of clause 41, wherein step d) is performed as a single or multiple steps, wherein the sample is correspondingly drawn to the other of the/each microfluidic channel corresponding to a reduced number of executions of force Or multiple consecutive positions.

43.一種對於一液體樣本進行一檢定之方法，該方法包含：a)將包含一或多個可壓縮氣體填充腔室之一微流控盒引入一讀數器裝置中，該讀數器裝置包含用於將該腔室壓縮/減壓之構件； b)向該微流控盒之一/該氣體填充腔室施加一力，以便將氣體之一部分從該腔室中排出；c)將一液體樣本引入該微流控盒中並且允許毛細管作用將該樣本抽吸至該微流控盒之一或多個微流控通道中，或減少施加至該氣體填充腔室之該力，以使得空氣被抽吸至該腔室中，導致一液體樣本被抽吸至該微流控通道中；d)減少施加至該微流控盒之該腔室之力，以使得空氣被抽吸至該腔室中，導致該液體樣本被進一步抽吸至該微流控通道中以便允許與一分析物結合劑及視需要存在於該通道中之一或多個進一步試劑接觸；e)允許在該微流控通道之一偵測區中形成並捕獲任何分析物/分析物結合劑複合物或分析物反應產物/分析物結合劑複合物；f)視需要增加施加至該微流控盒之該氣體填充腔室之力，以使得氣體從該腔室排出，導致液體從捕獲該分析物/分析物結合劑複合物之該微流控通道之至少一部分中排出，以使得所捕獲的分析物/分析物結合劑複合物存在於一實質上無液體環境中；及g)在該實質上無液體環境中偵測任何所捕獲分析物或分析物反應產物。 43. A method of performing a assay on a liquid sample, the method comprising: a) introducing a microfluidic cartridge containing one or more compressible gas-filled chambers into a reader device, the reader device comprising a member for compressing/decompressing the chamber; b) applying a force to one of the microfluidic cartridges/the gas-filled chamber to discharge a portion of the gas from the chamber; c) placing a liquid sample Introducing into the microfluidic cartridge and allowing capillary action to draw the sample into one or more microfluidic channels of the microfluidic cartridge, or reducing the force applied to the gas filled chamber to cause air to be Pumping into the chamber causes a liquid sample to be drawn into the microfluidic channel; d) reducing the force applied to the chamber of the microfluidic cartridge such that air is drawn into the chamber Causing the liquid sample to be further drawn into the microfluidic channel to allow contact with an analyte binding agent and optionally one or more further reagents in the channel; e) allowing for microfluidic control Form and capture any analyte/analysis in one of the channel detection zones a binder complex or an analyte reaction product/analyte binder complex; f) increasing the force applied to the gas-filled chamber of the microfluidic cartridge as needed to cause gas to escape from the chamber, resulting in liquid Dissipating at least a portion of the microfluidic channel capturing the analyte/analyte binder complex such that the captured analyte/analyte binder complex is present in a substantially liquid-free environment; and g) Any captured analyte or analyte reaction product is detected in this substantially liquid free environment.

44.如條款41或43之方法，其中所形成的該分析物/分析物結合劑複合物或分析物反應產物/分析物結合劑複合物包含磁性或順磁粒子。 44. The method of clause 41 or 43, wherein the analyte/analyte binder complex or analyte reaction product/analyte binder complex formed comprises magnetic or paramagnetic particles.

45.如條款44之方法，其中用於形成該等複合物之該等磁性粒子最初置放在與一磁鐵緊密接觸，或施加一磁力的該盒之表面相反的該微流控通道之一表面上，以使得該等磁性粒子被側向地抽吸穿過該微流控通道。 45. The method of clause 44, wherein the magnetic particles used to form the composite are initially placed in contact with a magnet, or a surface of the microfluidic channel opposite the surface of the cartridge to which a magnetic force is applied. Upper to cause the magnetic particles to be drawn laterally through the microfluidic channel.

46.如條款43-45之方法，其中步驟d)作為單一或多個步驟來執行，其中對應於力之減少執行次數，該樣本相應地被抽吸至該/每個微流控通道內之另一個或多個連續位置。 46. The method of clauses 43-45, wherein step d) is performed as a single or multiple steps, wherein the sample is correspondingly drawn into the/each microfluidic channel corresponding to a reduced number of executions of force Another or more consecutive locations.

47.如條款41-46中任一項之方法，其中從該腔室排出的導致從捕獲該分析物/分析物結合劑複合物之該微流控通道之至少一部分排出液體的氣體之體積足以導致該液體從該偵測區，但是不進一步沿著該微流控通道移除。 The method of any one of clauses 41 to 46, wherein a volume of gas discharged from the chamber that causes liquid to be discharged from at least a portion of the microfluidic channel that captures the analyte/analyte binder complex is sufficient This causes the liquid to pass from the detection zone but is not further removed along the microfluidic channel.

48.一種用於進行多重檢定，亦即複數個不同檢定的自給式拋棄式微流控盒，該微流控盒包含：一樣本輸入埠，該樣本輸入埠用於將一樣本引入該微流控盒及多個微流控通道中，該等微流控通道中之每一者適於接收該樣本之一部分並且能夠對於樣本之該部分進行一或多個檢定，以使得該微流控盒能夠偵測及/或判定該樣本中之多個不同分析物水準並且使用試劑來對於該樣本進行多個不同類型之檢定，該等試劑在樣本引入之前存在於該盒中。 48. A self-contained disposable microfluidic cartridge for performing multiple assays, that is, a plurality of different assays, the microfluidic cartridge comprising: the same input port, the sample input port is used to introduce the same book into the microfluidic device In the cartridge and the plurality of microfluidic channels, each of the microfluidic channels is adapted to receive a portion of the sample and to perform one or more assays on the portion of the sample to enable the microfluidic cartridge to A plurality of different analyte levels in the sample are detected and/or determined and reagents are used to perform a plurality of different types of assays on the sample, the reagents being present in the cassette prior to sample introduction.

49.如條款44之自給式拋棄式微流控盒，其用於如條款41-46中任一項之方法中。 49. The self-contained disposable microfluidic cartridge of clause 44, for use in the method of any of clauses 41-46.

50.如條款48之自給式拋棄式微流控盒，其進一步包含如條款1-26所定義之特徵。 50. The self-contained disposable microfluidic cartridge of clause 48, further comprising the features as defined in clauses 1-26.

51.如條款48-50中任一項之自給式拋棄式微流控盒，其能夠執行以下類型之檢定中之至少兩者、三者、四者、五者或更多者：免疫檢定、核酸檢定、基於受體的檢定、細胞計數檢定、比色檢定、酶檢定、電泳檢定、電化學檢定、光譜檢定、層析檢定、顯微鏡檢定、局部檢定、量熱檢定、濁度檢定、凝集檢定、黏度檢定、凝血檢定、凝結時間檢定、蛋白質合成 檢定、組織學檢定、培養檢定或滲透壓檢定。 51. The self-contained disposable microfluidic cartridge of any of clauses 48-50, which is capable of performing at least two, three, four, five or more of the following types of assays: immunoassay, nucleic acid Verification, receptor-based assay, cell count assay, colorimetric assay, enzyme assay, electrophoresis assay, electrochemical assay, spectral assay, chromatographic assay, microscopic assay, partial assay, calorimetric assay, turbidity assay, agglutination assay, Viscosity test, coagulation test, coagulation time test, protein synthesis test, histological test, culture test or osmotic pressure test.

52.如條款48-51中任一項之自給式拋棄式微流控盒，其能夠進行一組單獨檢定，該等檢定被設計來測試心臟病狀、腎上腺病狀、肝病狀、糖尿病或藥物濫用。 52. The self-contained disposable microfluidic cartridge of any of clauses 48-51, which is capable of performing a separate set of assays designed to test for heart disease, adrenal gland conditions, liver disease, diabetes or drug abuse .

53.如條款52之自給式拋棄式微流控盒，其用於偵測一心臟病狀並且其中該組單獨檢定用於偵測脂質水準、脂蛋白元；升半胱胺酸；C-反應蛋白(CRP)；及/或心臟酶。 53. The self-contained disposable microfluidic cartridge of clause 52, for detecting a heart condition and wherein the set is separately assayed for detecting lipid levels, lipoproteins; cysteine; C-reactive protein (CRP); and/or cardiac enzymes.

54.如條款52之自給式拋棄式微流控盒，其用於偵測一腎上腺病狀，並且其中該組單獨檢定用於偵測醛固酮、皮質醇、18-羥皮質醇及/或DHEA-S。 54. The self-contained disposable microfluidic cartridge of clause 52, for detecting an adrenal gland condition, and wherein the set of individual assays is for detecting aldosterone, cortisol, 18-hydroxycortisol, and/or DHEA-S .

55.如條款52之自給式拋棄式微流控盒，其用於偵測一肝臟病狀並且其中該組單獨檢定用於偵測一或多種肝臟酶、膽紅素、白蛋白、凝血酶原之一水準及/或一或多種病毒之存在。 55. The self-contained disposable microfluidic cartridge of clause 52, for detecting a liver condition and wherein the set of individual assays is for detecting one or more liver enzymes, bilirubin, albumin, prothrombin The presence of one level and/or one or more viruses.

56.如條款52之自給式拋棄式微流控盒，其用於偵測處於患上糖尿病之風險中之受試者或確認患有糖尿病之受試者並且其中該組單獨檢定用於偵測脂質水準、全血計數、禁食葡萄糖水準、血紅蛋白A1c及/或白蛋白。 56. The self-contained disposable microfluidic cartridge of clause 52, for detecting a subject at risk of developing diabetes or a subject confirmed to have diabetes and wherein the set is separately assayed for detecting lipid Level, whole blood count, fasting glucose level, hemoglobin A1c and/or albumin.

57.如條款52之自給式拋棄式微流控盒，其用於偵測藥物濫用，其中該組檢定用於偵測***；巴比妥酸鹽；丁基原啡因；苯二氮平；古柯鹼；搖頭丸；甲基***；***(鴉片劑/嗎啡)；***；三環抗抑鬱藥；及/或***。 57. The self-contained disposable microfluidic cartridge of clause 52, for detecting drug abuse, wherein the set of assays is for detecting amphetamine; barbiturate; butylmorphine; benzodiazepine; ***e Ecstasy; methamphetamine; heroin (opiates/morphine); methadone; tricyclic antidepressants; and/or cannabis.

58.一種用於進行多組檢定的多重檢定平臺，該多重檢定平臺包含如條款48-57中任一項之複數個微流控盒，每個盒能夠對於一樣本進行一組規定檢定及被構建來能夠接收及檢驗該等複數個微流控盒中之每一者的一讀數 器，其中該讀數器可被組配來偵測及/或判定可存在於該樣本中之一組分析物之水準。 58. A multi-validation platform for performing a plurality of sets of assays, the multi-assay platform comprising a plurality of microfluidic cartridges according to any one of clauses 48-57, each of which is capable of performing a set of prescribed assays and Constructing a reader capable of receiving and inspecting each of the plurality of microfluidic cartridges, wherein the reader can be configured to detect and/or determine a set of analytes that can be present in the sample The level.

59.如條款58之用於進行多組檢定之多重檢定平臺，其與如條款30-40中任一項之讀數器裝置一起使用。 59. A multiple assay platform for performing a plurality of sets of assays, as in clause 58, for use with a reader device according to any of clauses 30-40.

本發明現在藉由實例並且參照以下展示的圖式來進一步描述：圖1示出根據本發明之微流控盒；圖2詳細地示出如圖1識別之部分A；圖3示出根據本發明之讀數器；圖4示出圖3示出之讀數器之內部機構；圖5以平面圖來示出包含本發明之力控制構件的讀數器之內部部分；圖6示出沿著圖5之線A-A的截面圖；圖7：示出能夠每個盒運行不同數目之檢定的示範性盒格式之示意圖；圖8：示出根據本發明及Siemens Centaur C-肽檢定之偵測C-肽之比較繪圖。N=350；圖9：示出根據本發明相比於Siemens Centaur C-肽檢定之偵測C-肽之偏差繪圖比較。N=294；圖10：示出根據本發明及HemoIL D-二聚體HS500臨床分析器測試之偵測D-二聚體之比較繪圖；圖11：示出根據本發明及Siemens Dimension CRP臨床分析器測試之偵測CRP的比較繪圖； 圖12：示出根據本發明及Siemens Dimension hsCRP臨床分析器測試之偵測hsCRP的比較繪圖；圖13：示出加入血液中並且根據本發明來運作之惡性瘧原蟲(P.f)HRP2分析物之劑量響應曲線；圖14：示出用於多步肌鈣蛋白I檢定中之試劑之示意圖；圖15：示出涉及多步肌鈣蛋白I檢定中之步驟之圖示；圖16(a)及(b)：示出如與Siemens Centaur肌鈣蛋白Ultra測試相比，使用根據本發明之多步檢定在健康個體中量測之肌鈣蛋白I之繪圖；圖17示出在藉由空氣來移除緩衝液前後，根據本發明進行之C-肽檢定響應的比較；圖18示出在藉由空氣來移除液體樣本前後，根據本發明對於樣本血液進行之C-肽檢定響應之比較；及圖19示出使用本發明之盒的方法比較繪圖，該盒包含沒有氣體腔室之通道來控制液體填充及/或移除，以便執行INR測試及羅氏CoaguCheck INR測試。 The invention will now be further described by way of example and with reference to the accompanying drawings in which: FIG. 1 shows a microfluidic cartridge according to the invention; FIG. 2 shows in detail a portion A as identified in FIG. 1; Figure 4 shows the internal mechanism of the reader shown in Figure 3; Figure 5 shows the internal portion of the reader containing the force control member of the present invention in plan view; Figure 6 shows the same along Figure 5 Sectional view of line AA; Figure 7: Schematic diagram showing an exemplary cassette format capable of running a different number of assays per cartridge; Figure 8: shows detection of C-peptide according to the invention and the Siemens Centaur C-peptide assay Compare drawings. N = 350; Figure 9: shows a comparison of the deviations of the detected C-peptides compared to the Siemens Centaur C-peptide assay in accordance with the present invention. N=294; Figure 10: shows a comparative plot of the detected D-dimer according to the invention and the HemoIL D-dimer HS500 clinical analyzer test; Figure 11: shows the clinical analysis according to the invention and the Siemens Dimension CRP Comparative plot for detecting CRP in a test; Figure 12: Comparative plot showing detection of hsCRP in accordance with the present invention and the Siemens Dimension hsCRP clinical analyzer test; Figure 13: showing the malignancy added to the blood and operated in accordance with the present invention Dose response curve of Plasmodium (Pf) HRP2 analyte; Figure 14: Schematic diagram showing reagents used in multi-step troponin I assay; Figure 15: shows steps involved in multi-step troponin I assay Figure 16 (a) and (b): shows a plot of troponin I measured in healthy individuals using a multi-step assay in accordance with the present invention as compared to the Siemens Centaur troponin Ultra test; Figure 17 shows a comparison of the C-peptide assay response according to the present invention before and after the buffer is removed by air; Figure 18 shows the sample blood according to the present invention before and after the liquid sample is removed by air. Comparison of C-peptide assay responses; and Figure 19 A comparison is shown using a method of the cartridge of the present invention that includes a channel without a gas chamber to control liquid filling and/or removal for performing an INR test and a Roche CoaguCheck INR test.

圖1示出根據本發明之微流控盒(1)，其用於執行來自單一樣本之4個單獨檢定。盒(1)包含液體樣本輸入埠(3)，其連接至微流控通道(4)，該微流控通道劃分成複數個單獨通道(5及7)。每個通道(5)在盒(1)內延伸並且流體連接至氣體填充腔室(10)。不連接至氣體填充腔室之另一個通道(7)係用於多對照量測之對照通道。在使用中，流體樣本填充通道(5及7)並且此可藉由電極(未展示)來偵測到，該等電極與讀數器內之對應電氣觸點電 氣接觸。在讀數器偵測到樣本負載至盒(1)中之合適信號後，讀數器可開始檢定。亦提供用於接收液體之吸收器(13)。在吸收器之直接上游，存在液體止擋物(15)，其防止液體僅藉由毛細管作用來直接進入吸收器(13)中。因此，在最初使用毛細管施加來施加樣本時，液體樣本不經過液體止擋物(15)。 Figure 1 shows a microfluidic cartridge (1) according to the invention for performing 4 separate assays from a single sample. The cartridge (1) contains a liquid sample input port (3) that is coupled to a microfluidic channel (4) that is divided into a plurality of individual channels (5 and 7). Each channel (5) extends within the cartridge (1) and is fluidly connected to the gas filled chamber (10). The other channel (7) that is not connected to the gas-filled chamber is used for the control channel for multi-control measurements. In use, the fluid sample fills the channels (5 and 7) and this can be detected by electrodes (not shown) that are in electrical contact with corresponding electrical contacts in the reader. After the reader detects the appropriate load from the sample load to the cartridge (1), the reader can begin the assay. An absorber (13) for receiving liquid is also provided. Directly upstream of the absorber, there is a liquid stop (15) that prevents the liquid from entering the absorber (13) directly by capillary action only. Thus, the liquid sample does not pass through the liquid stop (15) when the sample is initially applied using capillary application.

更詳細地描述每個通道(5)，存在印刷特徵(20，22，24，26)，其被設計來在製造過程期間限制置放在每個通道(5)內之任何試劑之運動。與印刷特徵(20)相鄰並且藉由如圖2更詳細示出的部分A來表示的係較小尺寸通道(例如，0.1-0.2mm)(50)，其垂直地延伸遠離檢定通道(例如，0.75-1mm)(5)。在每個通道(50)內係單向閥(0.1mm乘以0.9mm)(52)，其被設計來在施加液體樣本時允許存在於每個通道(5)中之氣體或空氣退出盒(1)。因此，在藉由毛細管施加來將樣本施加至盒後，樣本填充通道(4)，將存在於通道(5)中之空氣移位，從而經由單向閥(52)退出盒。樣本藉由毛細管作用來填充直到樣本近似相鄰於每一側通道(50)為止。定位在印刷特徵(20)上方的係每個檢定通道(5)之第一反應區(28)，其中置放一或多個結合及/或反應劑，其被設計來與可存在於將要檢定之液體樣本中之特定分析物或其反應產物反應並結合。舉例而言，置放在該等通道(5)之第一區(28)中的可為磁性粒子，該等粒子用被設計來特異性結合將要偵測之分析物之第一表位的抗體來官能化。沉積在每個通道之第二區(30)中的可為螢光標記膠乳粒子，該等粒子用被設計來特異性結合將要偵測之分析物之不同表位之另一個抗體來官能化。定位在區域(28，30)之遠端/近端的係偵測區(32)，其中標記物/分析物/磁性粒子複合物可加以偵測。 Depicting each channel (5) in more detail, there are printed features (20, 22, 24, 26) that are designed to limit the movement of any reagent placed within each channel (5) during the manufacturing process. A smaller-sized channel (eg, 0.1-0.2 mm) (50) adjacent to the printed feature (20) and represented by portion A as shown in more detail in FIG. 2, extending vertically away from the assay channel (eg, , 0.75-1mm) (5). Within each channel (50) is a one-way valve (0.1 mm by 0.9 mm) (52) designed to allow gas or air present in each channel (5) to exit the cartridge when a liquid sample is applied ( 1). Thus, after application of the sample to the cartridge by capillary application, the sample fills the channel (4), displacing the air present in the channel (5), thereby exiting the cartridge via the one-way valve (52). The sample is filled by capillary action until the sample is approximately adjacent to each side channel (50). Positioned above the printed feature (20) as a first reaction zone (28) of each assay channel (5) in which one or more bonding and/or reactants are placed, which are designed to exist and are to be assayed The specific analyte or its reaction product in the liquid sample reacts and combines. For example, the first region (28) placed in the channels (5) may be magnetic particles that are designed to specifically bind to the first epitope of the analyte to be detected. To functionalize. The fluorescently labeled latex particles deposited in the second zone (30) of each channel are functionalized with another antibody designed to specifically bind to a different epitope of the analyte to be detected. A distal/proximal detection zone (32) positioned at the region (28, 30) wherein the marker/analyte/magnetic particle complex can be detected.

定位在偵測區(32)之遠端/近端的係氣體填充腔室(10)，其被 設計來與存在於本發明之讀數器裝置(如稍後描述)內之力施加特徵並置，以使得力施加特徵能夠向氣體填充腔室(10)施加力以便導致腔室(10)內之氣體從腔室(10)中排出並且進入檢定通道(5)中。施加至腔室(10)之力之減少導致空氣從檢定通道(5)中被抽吸回到腔室(10)中。 a gas-filled chamber (10) positioned at the distal/proximal end of the detection zone (32) that is designed to be juxtaposed with force application features present in the reader device of the present invention (as described later), The force application feature is enabled to apply a force to the gas filling chamber (10) to cause gas within the chamber (10) to exit the chamber (10) and enter the assay channel (5). The reduction in the force applied to the chamber (10) causes air to be drawn back into the chamber (10) from the assay channel (5).

在使用中，盒(1)***讀數器(100)中，如圖3示出。讀數器具有可關閉之門(102)，其可打開以便接近讀數器之盒接收埠(103)。一旦盒***讀數器(100)中並且將樣本施加至盒(1)，門(102)可關閉。讀數器容納多個特徵，其被設計來接觸盒(1)且/或促進執行本發明之檢定，如更詳細地描述。讀數器(100)之頂部表面包含觸控螢幕顯示器(104)，其允許使用者與讀數器(100)相互作用，以及接收關於任何檢定之效能的資訊。 In use, the cartridge (1) is inserted into the reader (100) as shown in FIG. The reader has a closable door (102) that can be opened to access the cassette of the reader to receive the cassette (103). Once the cartridge is inserted into the reader (100) and the sample is applied to the cartridge (1), the door (102) can be closed. The reader houses a plurality of features designed to contact the cartridge (1) and/or to facilitate performing the assay of the present invention, as described in more detail. The top surface of the reader (100) includes a touch screen display (104) that allows the user to interact with the reader (100) and receive information regarding the performance of any assay.

圖4示出讀數器(100)之內部特徵。讀數器包括可再充電蓄電池(110)，其用於為讀數器及其各種功能提供功率，如描述。為蓄電池(110)充電之功率經由DC插座(106)來提供。讀數器(100)進一步包括加熱器(111)，其用於在需要時將盒(1)加熱；光學器件區塊(112)，其含有用於偵測來自盒(1)之螢光信號的必需光學器件；可移動磁鐵(113)，其被設計來將磁性粒子固定在盒之偵測區(32)內；及杠桿機構(114)，其被設計來接觸盒(1)之腔室(10)並且施加力以便導致空氣從腔室(10)中排出。 Figure 4 shows the internal features of the reader (100). The reader includes a rechargeable battery (110) for providing power to the reader and its various functions, as described. The power to charge the battery (110) is provided via a DC outlet (106). The reader (100) further includes a heater (111) for heating the cartridge (1) when needed; an optics block (112) containing a fluorescent signal for detecting the cartridge (1) Necessary optics; movable magnet (113) designed to hold magnetic particles in the detection zone (32) of the cartridge; and lever mechanism (114) designed to contact the chamber of the cartridge (1) ( 10) and applying a force to cause air to escape from the chamber (10).

在使用中，盒(1)***讀數器(100)中直到盒接觸讀數器(100)內之對齊特徵(122)為止。盒(1)之正確***藉由存在於盒中之電極與存在於讀數器中之對應觸點來偵測。此情況向讀數器指示盒(1)已經正確***並且可開始檢定過程之啟動。指示電動機(120)啟用齒條-齒輪機構。齒輪(124)在順時針方向上轉動以便導致杠桿(128)之齒條機構(126)垂直地向上移動。此 運動導致杠桿(128)的呈指狀物形式之另一端(132)向下移動並且與盒(1)之腔室(10)接觸。電動機之持續運作導致齒條機構(126)向上，以及杠桿(128)之另一端(132)對應向下運動，以使得遞增之力施加至盒(1)之腔室(10)，從而將氣體從腔室(10)中排出。一旦所需量之氣體從腔室(10)中排出，杠桿(128)之末端(132)保持與氣體填充腔室(10)接觸以便防止氣體被抽吸回到腔室(10)中。在此時點，藉由顯示器(104)上之訊息通知使用者現在可將樣本施加至盒(1)。 In use, the cartridge (1) is inserted into the reader (100) until the cartridge contacts the alignment feature (122) within the reader (100). The correct insertion of the cartridge (1) is detected by the electrodes present in the cartridge and the corresponding contacts present in the reader. This condition indicates that the reader indicating box (1) has been properly inserted and the initiation of the verification process can begin. The indicator motor (120) enables the rack-and-gear mechanism. The gear (124) rotates in a clockwise direction to cause the rack mechanism (126) of the lever (128) to move vertically upward. This movement causes the other end (132) of the lever (128) in the form of a finger to move downward and into contact with the chamber (10) of the cartridge (1). The continued operation of the motor causes the rack mechanism (126) to move upward and the other end (132) of the lever (128) to move downwardly so that an increasing force is applied to the chamber (10) of the cartridge (1), thereby Discharged from the chamber (10). Once the desired amount of gas is expelled from the chamber (10), the end (132) of the lever (128) remains in contact with the gas filled chamber (10) to prevent gas from being drawn back into the chamber (10). At this point, the user is notified by the message on the display (104) that the sample can now be applied to the cartridge (1).

經由輸入埠(3)，樣本與盒(1)接觸並且引入其中。如前所述，樣本藉由毛細管作用來填充通道(4、5及7)，並且空氣經由閥(52)排放。在毛細管填充之後，在通道(5及7)中電性偵測到液體樣本之一部分，此指示讀數器可繼續運作。然後，引導電動機在逆時針方向上轉動齒輪機構(124)，進而導致齒條機構(126)在向下方向上並且杠桿(128)之另一端(132)向上，以使得施加至盒(1)之腔室(10)的力得以減少。如施加至腔室(10)之力之此減少導致空氣被抽吸回到腔室(10)中，進而將樣本抽吸至通道(5)之第一區(28)中。電動機(120)及相關聯杠桿運動能夠謹慎控制施加至腔室(10)之力之減少，從而控制液體樣本被抽吸至第一區(28)中之深淺程度。此亦可經由電極感測回饋來控制。進入通道(5)之第一區(28)中之液體樣本導致存在於第一區(28)中之功能衍生磁性粒子藉由樣本來再懸浮。電動機(120)停止一段時間以便允許可存在於液體樣本中之任何所需分析物結合至磁性粒子之表面上之功能分析物結合部分以便形成分析物/磁性粒子複合物。在一段規定時間之後，電動機再次啟動並且將力之進一步減少施加至腔室(10)，導致更多空氣被抽吸回到腔室(10)中，進而將樣本及分析物/磁性粒子複合物抽吸至通道(5)之第二區(30)中。每個通道(5)之第二區(30)含有功能衍生螢光標記膠乳粒 子，其能夠結合至分析物/磁性粒子複合物以便形成膠乳粒子/分析物/磁性粒子複合物層疊物。在另一段時間之後，施加至腔室(10)之力進一步減少並且存在於其中之液體及相關聯複合物被抽吸至偵測區(32)中。 Via the input 埠 (3), the sample is brought into contact with the cassette (1) and introduced therein. As previously mentioned, the sample fills the channels (4, 5 and 7) by capillary action and the air is discharged via the valve (52). After the capillary is filled, a portion of the liquid sample is electrically detected in the channels (5 and 7), which indicates that the reader can continue to operate. Then, the motor is guided to rotate the gear mechanism (124) in a counterclockwise direction, thereby causing the rack mechanism (126) to be in the downward direction and the other end (132) of the lever (128) to be upward so as to be applied to the cartridge (1) The force of the chamber (10) is reduced. This reduction in force, such as applied to the chamber (10), causes air to be drawn back into the chamber (10), thereby drawing the sample into the first zone (28) of the channel (5). The motor (120) and associated lever movements can carefully control the reduction in force applied to the chamber (10) to control how shallow the liquid sample is drawn into the first zone (28). This can also be controlled via electrode sensing feedback. The liquid sample entering the first zone (28) of the channel (5) causes the functionally-derived magnetic particles present in the first zone (28) to be resuspended by the sample. The motor (120) is stopped for a period of time to allow any desired analytes that may be present in the liquid sample to bind to the functional analyte binding portion on the surface of the magnetic particles to form an analyte/magnetic particle composite. After a specified period of time, the motor is started again and a further reduction in force is applied to the chamber (10), causing more air to be drawn back into the chamber (10), which in turn will sample and analyte/magnetic particle composites Pump into the second zone (30) of the channel (5). The second zone (30) of each channel (5) contains functionally derived fluorescently labeled latex particles that are capable of binding to the analyte/magnetic particle composite to form a latex particle/analyte/magnetic particle composite laminate. After a further period of time, the force applied to the chamber (10) is further reduced and the liquid and associated complex present therein are drawn into the detection zone (32).

一旦液體樣本及相關聯複合物被抽吸至偵測區(32)中，磁鐵(113)藉由電動機(150)及相關聯齒輪(152)及齒條(154)驅動以使得磁鐵與盒之偵測區(32)緊密鄰近，以使得藉由磁鐵(113)之磁力將磁性複合物吸引至磁鐵並且保持偵測區(32)內之適當位置中。其後，再應用電動機(120)以導致杠桿機構(114)增加施加至氣體填充腔室(10)之力，導致空氣再一次從腔室(10)中排出，導致將存在於偵測區(32)中之液體樣本及非磁性結合材料遠離偵測區(32)並且沿著通道(5)推動，並且液體之一部分退出至吸收器(13)中。可不需要將所有液體排出至吸收器(13)中並且事實上可僅需要將液體從偵測區(32)中移除，以使得所得磁性結合複合物存在於基本上空氣環境中。由於不像在側流產物中所發生的那樣使用額外樣本體積來執行清洗，並且不需要帶材上緩衝液囊或量計中緩衝液遞送系統，此可為尤其有利的。 Once the liquid sample and associated composite are pumped into the detection zone (32), the magnet (113) is driven by the motor (150) and associated gear (152) and rack (154) to cause the magnet and the cartridge The detection zone (32) is in close proximity so that the magnetic composite is attracted to the magnet by the magnetic force of the magnet (113) and remains in position within the detection zone (32). Thereafter, the motor (120) is applied again to cause the lever mechanism (114) to increase the force applied to the gas-filled chamber (10), causing the air to again exit the chamber (10), resulting in the presence of the detection zone ( The liquid sample and non-magnetic binding material in 32) are remote from the detection zone (32) and are pushed along the channel (5), and one of the liquids partially exits into the absorber (13). It may not be necessary to expel all of the liquid into the absorber (13) and in fact may only need to remove the liquid from the detection zone (32) such that the resulting magnetically bound composite is present in a substantially air environment. This may be particularly advantageous since the cleaning is performed using an additional sample volume as occurs in the lateral flow product, and does not require a buffer on the strip or a buffer delivery system in the meter.

電動機(120)能夠以可變速度來操作，因此將空氣抽吸至腔室(10)中及將空氣從腔室(10)中排出可容易地在不同速率下發生，並且存在於通道(5)及相關聯區(28、30及32)中之液體之流動速率係相應地可變的。 The electric motor (120) is capable of operating at a variable speed so that drawing air into the chamber (10) and expelling air from the chamber (10) can easily occur at different rates and are present in the passage (5) And the flow rate of the liquid in the associated zones (28, 30 and 32) is correspondingly variable.

在將液體從偵測區(32)移除之後，所捕獲的複合物存在於基本上無液體環境中並且可使用存在於光學區塊(112)中之偵測器來偵測。偵測器可呈例如分光光度計形式，其能夠偵測存在於捕獲膠乳粒子/分析物/磁性粒子複合物上之螢光標記物。 After the liquid is removed from the detection zone (32), the captured complex is present in a substantially liquid-free environment and can be detected using a detector present in the optical block (112). The detector can be in the form of, for example, a spectrophotometer capable of detecting fluorescent labels present on the captured latex particles/analyte/magnetic particle composite.

在關於圖4示出及描述之實施方式的替代實施方式中，壓電 彎曲機可用於控制施加至盒之氣體填充腔室之力。圖5示出力控制構件(200)。力控制構件(200)包含一系列壓電彎曲機(202)，其藉由固定區塊(204)固定在第一端(201)。每個壓電彎曲機亦在第一端處電氣地耦接至電氣連接(206)，該等連接控制提供至每個彎曲機(202)之電信號。如可發現，每個彎曲機(202)連接至其自己的一組電氣連接(206)，以使得每個彎曲機獨立地可控制。如圖6示出，每個彎曲機(202)之另一端(208)停留在底座(210)之頂部表面(209)上，該底座被設計來在使用中接觸本發明之微流控盒(220)之氣體腔室之外表面。 In an alternate embodiment of the embodiment shown and described with respect to Figure 4, a piezoelectric bending machine can be used to control the force applied to the gas filled chamber of the cartridge. Figure 5 shows the force control member (200). The force control member (200) includes a series of piezoelectric bending machines (202) that are secured to the first end (201) by a fixed block (204). Each piezoelectric bender is also electrically coupled at a first end to an electrical connection (206) that controls the electrical signals provided to each bender (202). As can be seen, each bender (202) is connected to its own set of electrical connections (206) such that each bender is independently controllable. As shown in Figure 6, the other end (208) of each bender (202) rests on a top surface (209) of the base (210) that is designed to contact the microfluidic cartridge of the present invention in use ( 220) The outer surface of the gas chamber.

圖6示出沿著圖5之線A-A之截面圖，以使得力控制構件(200)之各個部分及其如何起作用可更好地理解。在圖6中，力控制構件(200)與正確***讀數器中之微流控盒(220)一起展示以使得微流控盒之氣體填充腔室直接定位在力控制構件(200)之底座(210)下方。底座(210)之底部表面(212)經成形以接觸微流控盒(220)之氣體腔室之一部分並且經由藉由壓電彎曲機(202)施加至底座(210)之合適控制，底座(210)能夠將可變力施加至微流控盒(220)之氣體腔室。 Figure 6 shows a cross-sectional view along line A-A of Figure 5 so that the various portions of the force control member (200) and how they function can be better understood. In Figure 6, the force control member (200) is shown with the microfluidic cartridge (220) correctly inserted into the reader such that the gas filled chamber of the microfluidic cartridge is positioned directly at the base of the force control member (200) ( 210) below. The bottom surface (212) of the base (210) is shaped to contact a portion of the gas chamber of the microfluidic cartridge (220) and is suitably controlled via a piezoelectric bending machine (202) to the base (210), the base ( 210) A variable force can be applied to the gas chamber of the microfluidic cartridge (220).

如圖6示出，壓電彎曲機(202)在其未成形剛性狀態下。在此實施方式中，力控制構件(200)被構建來使得壓電彎曲機(202)能夠給予底座(210)最大力，以使得底座(210)之底部表面(212)向下推動並且壓縮氣體填充腔室，導致腔室內之氣體從腔室中排出。 As shown in Figure 6, the piezoelectric bending machine (202) is in its unformed rigid state. In this embodiment, the force control member (200) is constructed such that the piezoelectric bending machine (202) can impart maximum force to the base (210) such that the bottom surface (212) of the base (210) pushes down and compresses the gas Filling the chamber causes the gas in the chamber to exit the chamber.

雖然未展示，向壓電彎曲機(202)施加電荷導致壓電彎曲機(202)彎曲並且壓電彎曲機(202)之末端(208)向上彎曲。壓電彎曲機(202)之此向上彎曲減少施加至底座(210)之力，進而導致底座(210)減少施加至盒(220) 之氣體填充腔室之力。減少施加至氣體填充腔室之力提供氣體填充腔室之減壓及相應地氣體返回進入腔室中。經由合適電傳訊，可彎曲並放鬆壓電彎曲機(202)，導致氣體填充腔室相應地減壓或壓縮並且氣體排出或抽吸至腔室中。 Although not shown, applying a charge to the piezoelectric bending machine (202) causes the piezoelectric bending machine (202) to bend and the end (208) of the piezoelectric bending machine (202) to bend upward. This upward bending of the piezoelectric bender (202) reduces the force applied to the base (210), which in turn causes the base (210) to reduce the force applied to the gas filled chamber of the cartridge (220). Reducing the force applied to the gas-filled chamber provides a reduced pressure of the gas-filled chamber and correspondingly gas returning into the chamber. Via suitable electrical communication, the piezoelectric bender (202) can be bent and relaxed, causing the gas-filled chamber to be decompressed or compressed accordingly and the gas can be expelled or drawn into the chamber.

許多壓電彎曲機在此項技術中為已知的並且可適合用於本發明中。審閱本說明書之熟習此項技術者選擇適合於特定用途之彎曲機。本發明人使用具有多達幾毫米之位移及毫秒範圍內之響應時間的各種此等壓電彎曲機。電壓可程式化放大器可用於控制每個壓電彎曲機。合適放大器包括具有從50V至200V可程式化之全刻度輸出電壓的32-通道、14-位DAC(AD5535)或可從Analog Devices(Norwood,MA 02062,USA)獲得之高電壓四通道12-位電壓輸出DAC(AD5504)。可獲得1N-2N之力。 Many piezoelectric bending machines are known in the art and may be suitable for use in the present invention. Those skilled in the art who have reviewed this specification will select a bending machine that is suitable for a particular use. The inventors have used a variety of such piezoelectric bending machines having a displacement of up to several millimeters and a response time in the millisecond range. A voltage programmable amplifier can be used to control each piezoelectric bending machine. Suitable amplifiers include a 32-channel, 14-bit DAC (AD5535) with a programmable full-scale output voltage from 50V to 200V or a high-voltage four-channel 12-bit available from Analog Devices (Norwood, MA 02062, USA) Voltage Output DAC (AD5504). A force of 1N-2N can be obtained.

以上提供本發明之特定實施方式之描述，但是本發明被設計成可容易地調適之平臺形式。舉例而言，通氣孔位置可改變以允許毛細管填充至通道(5)內之不同位置，或通氣孔完全地省去並且樣本填充藉由將氣體從腔室(10)排出之後的主動填充來發生，並且藉由在釋放施加至腔室(10)之壓力之後空氣返回到氣體腔室中來將樣本抽吸至盒(1)及通道(5,7)中。 The above description of specific embodiments of the invention is provided, but the invention is designed in a form that can be easily adapted. For example, the vent position can be varied to allow the capillary to be filled into different locations within the channel (5), or the vent is completely omitted and the sample fill occurs by active filling after the gas is expelled from the chamber (10). And the sample is aspirated into the cartridge (1) and the channel (5, 7) by returning the air to the gas chamber after releasing the pressure applied to the chamber (10).

此外，讀數器可被設計成藉由產品要求所規定的一系列帶材大小來利用多種測試方式。帶材可被設計成針對特定產物組態及組測試而製造成例如2、4及10通道格式(參見圖5展示不同帶材大小)。不同帶材大小之可用性允許與目標市場中之確立產品相比，本發明系統以增加之效能及減少之成本結構來提供混合技術下之多重測試來滿足照護點市場之特定使用者要求。 In addition, the reader can be designed to utilize a variety of test methods by a range of strip sizes as specified by the product requirements. The strips can be designed to be fabricated into, for example, 2, 4, and 10 channel formats for specific product configurations and group tests (see Figure 5 for different strip sizes). The availability of different strip sizes allows the system of the present invention to provide multiple tests under hybrid technology to meet the specific user requirements of the point of care market with increased performance and reduced cost structure compared to established products in the target market.

參照展示不同尺寸帶材之圖7，2通道盒被設計成用於具有控制之單一檢定，4通道盒用於具有控制之2-3個分析物之組並且10通道盒允許執行混合技術及需要高多重處理能力之產品(例如，藥物濫用)之複雜檢定。所描述平臺具有非常靈活的樣本及檢定架構及讀數器控制及量測能力，允許在新的測試組或測試類型或照護點操作得以識別時，利用對於新機會的前向相容性。 Referring to Figure 7, which shows different sized strips, the 2-channel box is designed for a single assay with control, the 4-channel box is used for groups of 2-3 analytes with control and the 10-channel box allows for the implementation of mixing techniques and needs Complex verification of products with high multi-processing capabilities (eg, drug abuse). The described platform has a very flexible sample and verification architecture and reader control and measurement capabilities that allow for forward compatibility with new opportunities when new test groups or test types or care point operations are identified.

雖然主要量測技術係螢光，平臺亦併入電化學量測並且可容易地併入其他方法。此在以下進一步詳細地論述。 While the primary metrology technique is fluorescent, the platform incorporates electrochemical measurements and can be easily incorporated into other methods. This is discussed in further detail below.

為了在單一平臺上提供多種測試類型及方式，已開發一組靈活核心技術能力及控制，其可根據需要並且以提供不同檢定格式步驟的順序來使用。系統架構設計原則為： In order to provide multiple test types and methods on a single platform, a flexible set of core technology capabilities and controls have been developed that can be used as needed and in an order that provides different verification format steps. The system architecture design principles are:

■磁性粒子捕獲相 ■Magnetic particle capture phase

■液體運動控制 ■Liquid motion control

■液體從偵測區移除 ■The liquid is removed from the detection area

■空氣中之標記物檢測 ■Detection of markers in the air

■多通道多重處理 ■Multi-channel multiprocessing

■通道內多重處理 ■Multi-channel processing

■動態範圍 ■Dynamic range

■板上控制 ■Onboard control

■電化學量測 ■Electrochemical measurement

■加熱及溫度控制 ■ Heating and temperature control

■樣本預處理 ■ Sample pretreatment

此平臺架構允許在系統上安排許多不同測試類型及技術。每個技術核心原則在以下論述。 This platform architecture allows for many different test types and technologies to be arranged on the system. The core principles of each technology are discussed below.

磁性粒子捕獲及液體控制Magnetic particle capture and liquid control

已知使用粒子捕獲可改良捕獲動力學。對於免疫檢定，本發明之平臺使用順磁粒子作為捕獲表面。不同順磁粒徑可用於優化每個測試類型之效能。在檢定開發期間利用100至1000nm範圍內之順磁粒子。粒子捕獲相與螢光粒子標記相組合。同樣地，視檢定靈敏度及範圍要求而定，螢光粒子相之大小可改變。螢光粒子之典型大小可在40nm-4000nm範圍內。 It is known that particle capture can be used to improve capture kinetics. For immunoassays, the platform of the present invention uses paramagnetic particles as the capture surface. Different paramagnetic particle sizes can be used to optimize the performance of each test type. Paramagnetic particles in the range of 100 to 1000 nm are utilized during assay development. The particle capture phase is combined with the fluorescent particle label. Similarly, depending on the sensitivity and range requirements, the size of the fluorescent particle phase can vary. The typical size of the fluorescent particles can range from 40 nm to 4000 nm.

一些檢定，諸如C-反應蛋白(CRP)，需要量測相對高濃度之分析物並且利用與磁性粒子組合的直接螢光團標記抗體偶聯物，而高靈敏度檢定總體上利用與磁性粒子組合的螢光粒子標記。重要的是，樣本中之捕獲物及標記物相係可移動的以便驅動捕獲事件。帶材內之不當流動得以最小化的事實進一步有助於上述目標。在通道填充期間，樣本流動經過乾燥測試試劑。試劑溶解及因此流動前緣作用藉由使用允許良好通道填充但是導致受控較慢溶解的調配物來最小化。在最初樣本填充事件之後，流動停止以使得防止樣本進一步流動一段時間。此允許發生非常一致的溶解及隨後結合效率，因為沒有與基體相關的流動速率誤差影響所研究樣本體積或結合動力學。 Some assays, such as C-reactive protein (CRP), require the measurement of relatively high concentrations of analytes and direct labeling of antibody conjugates using direct fluorophores in combination with magnetic particles, while high sensitivity assays are generally utilized in combination with magnetic particles. Fluorescent particle markers. Importantly, the capture and marker phases in the sample are movable to drive capture events. The fact that the improper flow within the strip is minimized further contributes to the above objectives. During channel filling, the sample flows through the dry test reagent. Reagent solubilization and thus flow front effects are minimized by the use of formulations that allow for good channel filling but result in controlled slower dissolution. After the initial sample fill event, the flow stops so that the sample is prevented from flowing further for a period of time. This allows for very consistent dissolution and subsequent binding efficiency, as no flow rate error associated with the matrix affects the sample volume or binding kinetics studied.

與可變流動系統(例如，側流，Triage)相反，在視需要混合、靜態、固定體積中執行試劑溶解及分析物捕獲顯著改良檢定精確度及準確性。 In contrast to variable flow systems (eg, lateral flow, Triage), performing reagent solubilization and analyte capture in as needed mixed, static, fixed volumes significantly improves assay accuracy and accuracy.

在更複雜檢定，諸如肌鈣蛋白(如在別處描述)中，檢定作為 使用多個試劑區之多步程序來更有效地執行。在此情況下，能夠壓縮氣體腔室(10)以排出氣體並且執行從偵測區移除液體的量計功能亦用於實現盒(1)及相關聯通道內之精細液體運動控制。在樣本施加至盒(1)之前，氣體填充腔室(10)藉由量計壓縮，將氣體從腔室(10)及檢定通道排出。在樣本施加期間，腔室(10)藉由量計來保持壓縮並且樣本填充係藉由毛細管作用或完全在氣體驅動流體控制下。量計內之高解析度電動機或壓電彎曲機允許氣體腔室(10)上之壓力之非常受控的漸增釋放或增加，從而產生對於測試特定的壓力變化速率及量。此特徵提供多個重要優勢，包括使用任何壓電彎曲機之精細正性及負性彎曲來混合的能力。 In more complex assays, such as troponin (as described elsewhere), assays are performed more efficiently as a multi-step procedure using multiple reagent zones. In this case, the meter function capable of compressing the gas chamber (10) to vent gas and performing liquid removal from the detection zone is also used to achieve fine liquid motion control within the cartridge (1) and associated channels. Prior to application of the sample to the cartridge (1), the gas-filled chamber (10) is compressed by a gauge to expel gas from the chamber (10) and the assay channel. During sample application, the chamber (10) is kept compressed by a gauge and the sample is filled by capillary action or completely under gas-driven fluid control. A high resolution motor or piezoelectric bender within the meter allows for a very controlled incremental release or increase in pressure on the gas chamber (10), resulting in a specific rate and amount of pressure change for testing. This feature provides several important advantages, including the ability to mix using the fine positive and negative bends of any piezoelectric bender.

樣本填充時間可藉由引入試劑溶解、流體前緣作用及所研究樣本之體積的變化性而對於產品效能具有顯著影響。流體控制藉由直接控制樣本填充速率來減少填充時間之變化。流體控制允許樣本在受控時間移動的每個通道內之不同區，允許執行樣本預處理及多步檢定(本文描述)。流體控制及分離亦為NAT檢定所需要的封閉系統的必要條件(參見以下)。 The sample fill time can have a significant impact on product performance by introducing reagent lysis, fluid front effect, and volume variability of the sample under study. Fluid control reduces the change in fill time by directly controlling the sample fill rate. Fluid control allows for different zones within each channel of the sample to move at controlled times, allowing for sample pre-processing and multi-step verification (described herein). Fluid control and separation are also necessary for a closed system required for NAT verification (see below).

從偵測區移除液體Remove liquid from the detection zone

達成液體運動及控制的方法係使用電動機及力施加器，或接觸流體腔室(10)之壓電彎曲機機構來壓縮或釋放測試盒上之氣體腔室(10)。來自每個腔室(10)之所得氣體運動允許精細控制樣本及試劑之運動，包括將未結合標記物從測試通道之偵測區(32)中移除及視需要進入吸收器區域(13)中。 The method of achieving liquid motion and control is to compress or release the gas chamber (10) on the test cartridge using a motor and force applicator, or a piezoelectric bender mechanism that contacts the fluid chamber (10). The resulting gas motion from each chamber (10) allows fine control of movement of the sample and reagents, including removal of unbound label from the detection zone (32) of the test channel and, if desired, into the absorber region (13) in.

每個盒內之嵌入流體控制功能帶來許多重要差異化優勢。 The embedded fluid control function in each box brings many important differentiation advantages.

首先，所描述系統使用液體運動之氣體控制來提供結合及未 結合檢定組分之很有效分離。此係重要的，因為其完全避免帶材上液體試劑囊或量計中可更換液體清洗試劑包之複雜性及成本。 First, the described system uses gas motion control of liquid motion to provide a very efficient separation of the bound and unbound assay components. This is important because it completely avoids the complexity and cost of a liquid reagent cartridge on a strip or a replaceable liquid wash kit in a meter.

其次，本發明進一步使得能夠使用具有很低盒成本之夾層物製造技術及使用高通量、高控制捲筒生產系統的可製造性。 Second, the present invention further enables the use of sandwich manufacturing techniques with very low box costs and the manufacturability of high throughput, high control reel production systems.

第三，藉由使用氣體來將樣本及未結合標記物從偵測區(32)中移除意味著螢光標記之量測可在基本上無液體、氣體環境中進行。 Third, the removal of the sample and unbound label from the detection zone (32) by the use of a gas means that the measurement of the fluorescent marker can be carried out in a substantially liquid-free, gaseous environment.

空氣中之標記物檢測Marker detection in the air

與先前技術產品之標準檢定協定相比，氣體中之標記物量測產生進行螢光量測之多個顯著技術優勢。 Marker measurements in gases produce a number of significant technical advantages for performing fluorescence measurements compared to standard assay protocols for prior art products.

使用基本上氣體環境顯著減少液體樣本之淬滅效應，從而移除檢定變化及基體效應之主要來源。舉例而言，血細胞及血漿蛋白質之存在淬滅螢光信號，減少靈敏度並且增加螢光量測之變化性。在氣體或空氣環境中量測螢光團使得能夠使用螢光團，由於樣本淬滅，其不一定會被選擇。此允許更簡單光學設計、針對每個檢定來優化螢光團及單一通道內之多重處理。如以下藉由實例並且參照圖15及16來描述，與在緩衝液或全血中偵測相比，空氣中之偵測提供靈敏度之顯著改良。 The use of a substantially gaseous environment significantly reduces the quenching effect of the liquid sample, thereby removing the primary source of assay variation and matrix effects. For example, the presence of blood cells and plasma proteins quenches the fluorescent signal, reducing sensitivity and increasing variability in fluorescence measurements. Measuring the fluorophore in a gas or air environment enables the use of fluorophores, which are not necessarily selected due to sample quenching. This allows for a simpler optical design, optimized for the fluorophore and multiple processing within a single channel for each assay. As described below by way of example and with reference to Figures 15 and 16, detection in air provides a significant improvement in sensitivity compared to detection in buffer or whole blood.

總之，使用氣體來移除樣本及未結合標記物方法藉由減少樣本基體淬滅作用來減少檢定變化並且准許利用更大範圍之螢光團來用於檢定優化。此轉化成檢定設計靈活性、檢定速度及無與倫比的效能。 In summary, the use of gas to remove sample and unbound label methods reduces assay variation by reducing sample matrix quenching and permits the use of a wider range of fluorophores for assay optimization. This translates into verification design flexibility, speed of verification, and unparalleled performance.

多通道多重處理Multi-channel multiprocessing

本發明之平臺具有多通道及通道內多重處理能力。組測試可經由單一帶材內之多個通道以及量測每個通道中之標記物，例如螢光強度 的掃描光學頭來提供。通道之數目可視產品要求來變化。 The platform of the present invention has multiple channels and multiple processing capabilities within the channel. Group testing can be provided via multiple channels within a single strip and by measuring the markers in each channel, such as a fluorescent intensity scanning optics. The number of channels can vary depending on product requirements.

此允許開發其中每個通道含有不同檢定之組測試例如心臟組、代謝組等。因為個別檢定在獨立通道中係空間不同的，所以在多通道帶材中，每個檢定可以獨特試劑來組配。此帶來許多關鍵優勢：首先，每個檢定可針對以下目的來使用包括試劑、緩衝劑、pH等之最佳調配物：試劑之溶解、抗凝血、基體效應(HAMA等)之中和、最佳靈敏度、線性、範圍及檢定之穩定性。不需要為了開發組產品來發現多個檢定試劑組之相容優化或折衷檢定效能。每個檢定可存在於個別通道內之其自己最佳調配物中並且保持其相應高檢定效能。 This allows the development of group tests in which each channel contains different assays such as heart groups, metabolomes, and the like. Because individual assays have different spatial spaces in separate channels, each assay can be combined with a unique reagent in a multi-channel strip. This brings many key advantages: First, each assay can use the best formulation including reagents, buffers, pH, etc. for the following purposes: reagent dissolution, anticoagulant, matrix effect (HAMA, etc.) neutralization, Optimal sensitivity, linearity, range and stability of the assay. There is no need to discover compatible optimization or compromise assay performance for multiple assay reagent sets in order to develop a set of products. Each assay can be present in its own optimal formulation within an individual channel and maintain its corresponding high assay performance.

相反，單一通道內之多重測試固有地折衷個別測試之效能，因為試劑調配物必須與所有檢定相容。個別檢定要求經常有衝突，例如像pH那樣的基本項目顯著影響檢定效能。 In contrast, multiple tests within a single channel inherently compromise the performance of individual tests because the reagent formulation must be compatible with all assays. Individual verification requirements often conflict, for example, basic items like pH significantly affect verification performance.

多通道多重處理轉化成組測試設計靈活性、簡單性及組檢定開發速度及在各個組中保持單一檢定效能。 Multi-channel multiprocessing translates into group test design flexibility, simplicity, and group verification development speed and maintains a single verification performance across groups.

其次，多通道方法允許本發明平臺實現將不同檢定技術及不同轉換方法在單一帶材上組合的新穎組產品。 Second, the multi-channel approach allows the platform of the present invention to implement novel sets of products that combine different assay techniques and different conversion methods on a single strip.

越來越多的證據表明分子家族之量測可優於此家族之單一分子的量測。舉例而言，用於充血性心力衰竭分級中之利尿鈉肽總體上分離成BNP及NT-proBNP測試。多通道多重處理允許在一個帶材上量測proBNP、BNP、NT-proBNP及其他利尿鈉肽形式並且避免肽家族內之抗體表位交叉。相反，通道內多重處理導致分子家族量測之非特異性增加。目前描述之多通道方法適用於肌鈣蛋白測試市場，其中不同肌鈣蛋白同功異型 物可在單獨通道中量測以改良心肌梗死之診斷。 There is increasing evidence that the measurement of molecular families can be superior to the measurement of a single molecule of this family. For example, natriuretic peptides used in the classification of congestive heart failure are generally isolated as BNP and NT-proBNP tests. Multi-channel multiplex processing allows the measurement of proBNP, BNP, NT-proBNP and other natriuretic peptide forms on one strip and avoids antibody epitope crossover within the peptide family. In contrast, multiple processing within the channel results in a non-specific increase in molecular family measurements. The multi-channel approach described so far is applicable to the troponin test market, where different troponin isoforms can be measured in separate channels to improve the diagnosis of myocardial infarction.

通道內多重處理Multiple processing within the channel

當需要比率量測時，例如HbA1c及血液離子量測，為了達成最精確檢定效能，通道內多重處理係必不可少的。本發明平臺藉由在單一通道內量測一個以上螢光團來達成此。 When ratiometric measurements are required, such as HbA1c and blood ion measurements, multiple processing within the channel is essential to achieve the most accurate assay performance. The platform of the present invention accomplishes this by measuring more than one fluorophore in a single channel.

多通道及通道內多重處理之組合允許具有改良精確度及置信度之板上控制的靈活及強大的產品組合。 The combination of multi-channel and multi-channel processing allows for a flexible and powerful portfolio of on-board controls with improved accuracy and confidence.

動態範圍Dynamic Range

將要量測之分析物之較大動態範圍可經常為檢定效能之限制。舉例而言，肌鈣蛋白測試需要非常靈敏但是同時必須能夠量測高濃度以便監測在心肌梗塞患者中觀察到之變化。動態範圍經常在所需可量測範圍中產生非線性，影響精確度及準確性。 The large dynamic range of the analyte to be measured can often be a limiting factor in assay performance. For example, troponin testing needs to be very sensitive but at the same time must be able to measure high concentrations in order to monitor changes observed in patients with myocardial infarction. Dynamic range often produces nonlinearities in the required measurable range, affecting accuracy and accuracy.

多通道設計允許將具有較大動態範圍之挑戰性測試劃分成帶材上之多個通道，從而以線性方式來涵蓋所需可量測範圍之高靈敏度及高濃度。 The multi-channel design allows the challenging test with a large dynamic range to be divided into multiple channels on the strip to cover the high sensitivity and high concentration of the desired measurable range in a linear fashion.

對於肌鈣蛋白(I及/或T形式)，一個通道可含有對於0-100pg/ml之量測來優化之試劑，同時另一個通道可含有經優化來量測50-1000pg/ml之試劑並且另一個通道針對500-50000pg/ml來優化。靈敏度及範圍各自具有其自己的校準參數，並且樣本濃度從兩個結果之置信區間來指派。 For troponin (I and/or T forms), one channel may contain reagents optimized for a measurement of 0-100 pg/ml, while another channel may contain reagents optimized to measure 50-1000 pg/ml and The other channel is optimized for 500-50000 pg/ml. Sensitivity and range each have their own calibration parameters, and the sample concentration is assigned from the confidence intervals of the two results.

板上控制Onboard control

本發明平臺併入板上控制特徵來檢驗所獲得之測試結果之 有效性。每個測試類型需要獨特板上檢定控制以及多個通用特徵。所有測試可具有填充偵測以確保恰當樣本施加並且已用盒不能再測試。在需要時，盒併入血球比容量測結果以便調整受血球比容變化影響之彼等測試。可實行特定通道控制以併入低及高控制，其用於校準其餘血液基體變數及/或獨立地檢驗測試結果。耗盡控制可用於檢查人類抗小鼠抗體(HAMA)或其他樣本依賴性變數。 The platform of the present invention incorporates on-board control features to verify the validity of the test results obtained. Each test type requires unique on-board verification control and multiple common features. All tests can have fill detection to ensure proper sample application and the box can no longer be tested. When needed, the cartridge incorporates blood cell specific volume measurements to adjust for the effects of hematocrit changes. Specific channel control can be implemented to incorporate low and high controls for calibrating the remaining blood matrix variables and/or independently verifying the test results. Depletion control can be used to examine human anti-mouse antibodies (HAMA) or other sample dependent variables.

微處理器及相關聯軟體可控制每個特定檢定之定時、溫度、流體控制等，因為此等在單一盒內可具有不同要求。 The microprocessor and associated software can control the timing, temperature, fluid control, etc. of each particular assay, as these can have different requirements within a single cartridge.

電化學量測Electrochemical measurement

雖然所描述主要偵測方法係螢光，但是可進行其他光學量測及/或亦可在本發明平臺上進行電化學量測以併入傳統電化學測試方式(例如，葡萄糖測試)。另外，可對於同一帶材進行電化學及螢光量測，例如，具有C-肽螢光免疫檢定與電化學葡萄糖量測之糖尿病組。離子及血液氣體之習知離子選擇性電極(ISE)量測方法亦可用於本發明平臺。諸如螢光之光學與電化學轉換技術之組合使得能夠提供多種不同組測試。 Although the primary detection method described is fluorescent, other optical measurements can be performed and/or electrochemical measurements can also be performed on the platform of the present invention to incorporate conventional electrochemical test methods (eg, glucose testing). In addition, electrochemical and fluorescent measurements can be performed on the same strip, for example, a diabetic group with C-peptide fluorescent immunoassay and electrochemical glucose measurement. A conventional ion selective electrode (ISE) measurement method for ions and blood gases can also be used in the platform of the present invention. The combination of optical and electrochemical conversion techniques such as fluorescence enables a wide variety of different sets of tests to be provided.

加熱及溫度控制Heating and temperature control

溫度係大多數測試中之重要變數。對於一些檢定，溫度效應可使用溫度校正演算法來補償。然而，此對於判定個別盒批次經常係成問題的並且固定補償本身可變成誤差來源。在所有過程及基體變數中對溫度概況進行表徵可顯著影響產品之開發週期。在一些產品諸如PT/INR及分子測試中，適當溫度控制對於測試之功能及效能係關鍵的。本發明平臺允許併入整合加熱能力，其為每個測試類型提供最佳溫度要求。典型操作溫度 用於免疫檢定(34℃)、PT/INR(37℃)及核酸偵測(>37℃)等。加熱能力可優化提供帶材及預處理控制溫度範圍以利於測試協定中之最大靈活性。 Temperature is an important variable in most tests. For some assays, temperature effects can be compensated using a temperature correction algorithm. However, this is often problematic for determining individual cartridge batches and the fixed compensation itself can become a source of error. Characterizing the temperature profile across all process and matrix variables can significantly affect the product development cycle. In some products such as PT/INR and molecular testing, proper temperature control is critical to the function and performance of the test. The platform of the present invention allows for the incorporation of integrated heating capabilities that provide optimal temperature requirements for each test type. Typical operating temperatures are used for immunoassays (34 ° C), PT / INR (37 ° C) and nucleic acid detection (> 37 ° C). The heating capacity optimizes the strip and pre-treatment control temperature range to maximize flexibility in the test protocol.

樣本預處理Sample pretreatment

帶材上樣本運動之控制允許在將樣本提供至檢定特定試劑之前進行樣本預處理。此方法可應用於免疫檢定以便例如移除干擾物諸如HAMA物質或應用於脂質組以便移除特定脂質量測之不當組分(例如，HDL)。帶材上流體步驟模擬藉由臨床分析器使用的優化產物效能的能力，從而允許在產物開發期間樣本基體及干擾物得以快速分解。 Control of sample motion on the strip allows for sample pre-treatment prior to providing the sample to the assay specific reagent. This method can be applied to immunoassays to, for example, remove interfering substances such as HAMA species or to apply to lipid groups in order to remove inappropriate components (eg, HDL) of a particular lipid mass. The fluid step on the strip simulates the ability of the clinical analyzer to optimize product performance, allowing rapid decomposition of the sample matrix and interferents during product development.

示例性測試描述及測試資料Exemplary test description and test data

一步免疫檢定 One-step immunoassay

概述測試順序： Outline the test sequence:

1.盒***讀數器中 1. Insert the cartridge into the reader

2.盒氣體腔室藉由讀數器來壓縮 2. The cartridge gas chamber is compressed by the reader

3.藉由毛細管作用來填充或藉由讀數器控制填充，將樣本施加至盒。 3. Apply the sample to the cartridge by capillary action or by controlling the fill by a reader.

4.盒填充偵測電極濕化判定測試開始時序 4. Box fill detection electrode humidification determination test start timing

5.樣本將乾燥試劑再水化，該等乾燥試劑含有： 5. The sample rehydrates the dried reagents, which contain:

a.抗分析物抗體(表位1)官能化順磁粒子相 a. Anti-analyte antibody (epitope 1) functionalized paramagnetic particle phase

b.抗分析物抗體(表位2)官能化螢光標記物/粒子相 b. Anti-analyte antibody (epitope 2) functionalized fluorescent label / particle phase

6.試劑混合並且結合包含在樣本中之分析物，形成免疫檢定層疊物複合物(螢光標記物/粒子-分析物-順磁粒子)。 6. The reagent is mixed and combined with the analyte contained in the sample to form an immunoassay stack complex (fluorescent label/particle-analyte-paramagnetic particle).

7.結合反應發生歷時規定時間量(通常2分鐘)。 7. The binding reaction takes place over a specified amount of time (usually 2 minutes).

8.將磁場施加至帶材，定域至光學偵測區，將順磁粒子積聚至此位 置，在每個通道中形成粒子-分析物-標記物複合物條帶。 8. Apply a magnetic field to the strip, localize to the optical detection zone, accumulate paramagnetic particles to this location, and form a particle-analyte-label complex strip in each channel.

9.然後，液體樣本及未結合標記物移除步驟藉由讀數器開始將力施加至盒氣體腔室來執行。此壓縮力將氣體從氣體腔室經由測試通道排出，導致樣本液體及未結合螢光標記物/粒子從偵測區及視需要通道中排出並且進入樣本廢物吸收器中。在整個此步驟中施加磁場，藉由磁場將順磁粒子-分析物-標記物複合物保持在偵測區位置，同時將樣本從此區域排出。 9. The liquid sample and unbound label removal steps are then performed by the reader beginning to apply a force to the cartridge gas chamber. This compressive force discharges gas from the gas chamber through the test channel, causing the sample liquid and unbound fluorescent markers/particles to exit the detection zone and the desired channel and into the sample waste absorber. A magnetic field is applied throughout this step, and the paramagnetic particle-analyte-label complex is held in the detection zone position by the magnetic field while the sample is discharged from this zone.

10.量計光學頭在整個帶材上掃描並且量測每個通道之螢光強度。螢光強度與分析物濃度成比例。每個帶材批次及分析物通道個別地校準以使得螢光強度轉換成分析物濃度。 10. The metering optical head scans the entire strip and measures the intensity of the fluorescence of each channel. The fluorescence intensity is proportional to the analyte concentration. Each strip batch and analyte channel are individually calibrated to convert the fluorescence intensity to an analyte concentration.

一步免疫檢定之示例性效能資料集在圖8-13示出： An exemplary efficacy data set for a one-step immunoassay is shown in Figures 8-13:

C-肽 C-peptide

C-肽係在前胰島素分子中將胰島素之A-鏈連接至其B-鏈的短31-胺基酸多肽。前胰島素裂解成等莫耳濃度之胰島素及C-肽。在診斷情形下，C-肽用作胰島素之替代生物標記物並且用於監測糖尿病患者中之β-細胞功能(胰島素產生)。本發明人進行本發明檢定相對於商購ADVIA Siemens Centaur檯面系統之比較(參見圖8)。 The C-peptide is a short 31-amino acid polypeptide that links the A-chain of insulin to its B-chain in a proinsulin molecule. The pro-insulin is cleaved into insulin and C-peptide at a molar concentration. In the case of diagnosis, C-peptides are used as surrogate biomarkers for insulin and are used to monitor β-cell function (insulin production) in diabetic patients. The inventors performed a comparison of the assay of the invention with respect to the commercially available ADVIA Siemens Centaur countertop system (see Figure 8).

下表1展示對於所指示C-肽範圍，在參考系統之給定偏差內的結果之百分比。此展示本發明檢定達成在參考系統之20%內的通常約95%之結果。 Table 1 below shows the percentage of results within a given deviation of the reference system for the indicated C-peptide range. This demonstrates that the assay of the invention achieves typically about 95% of the results within 20% of the reference system.

對於超過0.5ng/ml之樣本(294個點)，執行本發明系統相比於Siemens Centaur參考系統之偏差分析，此在(圖9)中與確立商購臨床分析器相比來繪圖。每個點與參考系統值之偏差百分比相比於參考值來繪圖。繪圖示出本發明檢定系統具有與確立實驗室系統可比較的臨床準確性。 For samples over 0.5 ng/ml (294 points), a bias analysis of the system of the invention compared to the Siemens Centaur reference system was performed, which was plotted in (Fig. 9) compared to establishing a commercial clinical analyzer. The percentage deviation of each point from the reference system value is plotted against the reference value. The plot shows that the assay system of the present invention has clinical accuracy comparable to established laboratory systems.

D-二聚體 D-dimer

D-二聚體係纖維蛋白降解產物(FDP)，亦即在血塊藉由血纖維蛋白溶解來降解之後存在於血液中的較小蛋白質片段。D-二聚體分子含有纖維蛋白蛋白質之兩個交聯D片段。 D-dimerization system fibrin degradation products (FDP), that is, smaller protein fragments present in the blood after degradation of the clot by fibrinolysis. The D-dimer molecule contains two cross-linked D fragments of fibrin protein.

D-二聚體濃度用於幫助診斷血栓形成。其係在患有疑似血栓性病症之患者中執行之重要測試。雖然陰性結果實際上排除血栓形成，但是陽性結果可指示血栓形成，但是不排除其他可能原因。因此，其主要用途係排除血栓栓塞病，其中概率較低。 D-dimer concentrations are used to help diagnose thrombosis. It is an important test performed in patients with suspected thrombotic disorders. Although a negative result actually excludes thrombosis, a positive result may indicate thrombosis, but other possible causes are not excluded. Therefore, its main use is to exclude thromboembolic disease, in which the probability is low.

發明人使用目前描述之方法來執行劑量響應分析並且將結果與來自HemoIL D-二聚體HS 500(商購臨床分析器)之結果比較(參見圖10)。 The inventors performed the dose response analysis using the methods currently described and compared the results to results from HemoIL D-dimer HS 500 (commercially available clinical analyzer) (see Figure 10).

C-反應蛋白(CRP) C-reactive protein (CRP)

C-反應蛋白(CRP)係存在於血漿中之環形(環狀)、五聚體蛋白質，其水準響應於炎症而升高。其係來源於肝之急性期蛋白質，其在藉由 巨噬細胞及T細胞分泌介白素-6之後增加。 C-reactive protein (CRP) is a circular (cyclic), pentameric protein present in plasma whose level is elevated in response to inflammation. It is derived from the acute phase protein of the liver, which is increased by the secretion of interleukin-6 by macrophages and T cells.

CRP對於許多疾病類型具有診斷效用，該等疾病類型可如下匯總： CRP has diagnostic utility for many disease types, which can be summarized as follows:

1.1型糖尿病患者中之炎症狀態 Inflammatory state in patients with type 1 diabetes

2.用於感染控制及一般感染狀態之抗生素管理 2. Antibiotic management for infection control and general infection status

3.心血管疾病 3. Cardiovascular disease

4.某些癌症 4. Certain cancers

方法比較繪圖在圖11示出。所需可報告範圍係5-200μg/ml。 A method comparison plot is shown in FIG. The required reportable range is 5-200 μg/ml.

高靈敏度CRP(hs-CRP) High sensitivity CRP (hs-CRP)

高靈敏度CRP(hs-CRP)用於評估患上心血管疾病之風險。一般準則如下： High sensitivity CRP (hs-CRP) is used to assess the risk of developing cardiovascular disease. The general guidelines are as follows:

1.低：低於1.0mg/L之hs-CRP水準 1. Low: hs-CRP level below 1.0mg/L

2.平均值：在1.0與3.0mg/L之間 2. Average: between 1.0 and 3.0 mg/L

3.高：高於3.0mg/L 3. High: higher than 3.0mg/L

方法比較繪圖在圖12示出。資料證明本發明平臺完全能夠量測所需濃度之hs-CRP。 A method comparison plot is shown in FIG. The data demonstrates that the platform of the present invention is fully capable of measuring the desired concentration of hs-CRP.

瘧疾惡性瘧原蟲HRP2 Plague Plasmodium falciparum HRP2

瘧疾寄生蟲惡性瘧原蟲分泌富含組胺酸之蛋白質II(HRP2)，其用作生物標記物以偵測瘧疾寄生蟲惡性瘧原蟲(Pf)之存在。本發明平臺用於證明血液樣本中之HRP2之量測。將HRP2蛋白質加入血液中並且在本發明平臺及標準診斷(SD)瘧疾Pf快速測試中量測。 The malaria parasite Plasmodium falciparum secretes histidine-rich protein II (HRP2), which acts as a biomarker to detect the presence of the malaria parasite Plasmodium falciparum (Pf). The platform of the present invention is used to demonstrate the measurement of HRP2 in a blood sample. HRP2 protein was added to the blood and measured in the platform of the invention and the standard diagnostic (SD) malaria Pf rapid test.

本發明平臺上所量測的最低HRP2濃度係0.25ng/ml。相比之 下，使用SD測試，觀察到5ng/ml之很淡條帶。更低濃度係不可量測的。相對於SD測試量測5ng/ml濃度需要推薦30分鐘測試時間，0.25ng/ml本發明平臺測試結果耗費7分鐘。競爭測試需要30min檢定時間來洗掉未結合金溶膠標記物及任何裂解血液以便溶解很低濃度。亦存在向帶材施加緩衝液以便執行此清洗步驟的額外使用者操作。 The lowest HRP2 concentration measured on the platform of the invention was 0.25 ng/ml. In comparison, using the SD test, a very light band of 5 ng/ml was observed. Lower concentrations are not measurable. A 30-minute test time is recommended for a 5 ng/ml concentration relative to the SD test, and a 0.25 ng/ml platform test result of the invention takes 7 minutes. The competition test requires a 30 min assay time to wash away the unbound gold sol label and any lysed blood to dissolve very low concentrations. There are also additional user operations that apply a buffer to the strip to perform this cleaning step.

將資料分析並且結果在圖13中匯總。與具有快得多測試時間之SD測試相比，本發明檢定能夠量測顯著更低HRP2濃度。此檢定具有滿足快速測試以監測人群瘧疾根除計劃中之殘留感染之要求的靈敏度。 The data was analyzed and the results are summarized in Figure 13. The assay of the invention is capable of measuring significantly lower HRP2 concentrations than SD assays with much faster test times. This test has the sensitivity to meet the requirements of rapid testing to monitor residual infections in the population's malaria eradication program.

多步免疫檢定-例如肌鈣蛋白 Multi-step immunoassay - for example, troponin

本發明平臺可組配來執行多步檢定，允許發生逐步結合反應以優化結合動力學、測試時間及靈敏度。 The platform of the present invention can be configured to perform multi-step assays, allowing for a stepwise binding reaction to optimize binding kinetics, test time, and sensitivity.

在高靈敏度肌鈣蛋白檢定中，對於很低濃度之肌鈣蛋白，將抗體順磁粒子結合步驟與標記物/粒子結合步驟分離以顯著改良分析物-抗體順磁粒子結合步驟之結合速率及捕獲效率。隨後分別使用高親和力抗異硫氰酸螢光素及生物素-鏈黴親和素官能化粒子來逐步結合標記物粒子及順磁粒子實現更高捕獲及經結合肌鈣蛋白複合物之轉換。 In the high-sensitivity troponin assay, for very low concentrations of troponin, the antibody paramagnetic particle binding step is separated from the label/particle binding step to significantly improve the binding rate and capture of the analyte-antibody paramagnetic particle binding step. effectiveness. High affinity anti-isothiocyanate luciferin and biotin-streptavidin functionalized particles are then used to gradually bind the label particles and paramagnetic particles to achieve higher capture and conversion of the bound troponin complex.

概述測試順序： Outline the test sequence:

1.盒***讀數器中 1. Insert the cartridge into the reader

2.氣體腔室藉由讀數器來壓縮 2. The gas chamber is compressed by a reader

3.將樣本施加至盒，藉由毛細管作用來填充至第一通氣孔-止擋特徵，其中定位第一試劑(加標記之抗體) 3. Apply the sample to the cartridge and fill it by capillary action to the first vent-stop feature, wherein the first reagent (labeled antibody) is positioned

4.試劑再溶解及抗體-分析物培育及結合時間。 4. Re-dissolution of the reagent and antibody-analyte incubation and binding time.

5.較小腔室減壓導致液體樣本被進一步沿著通道抽吸，將樣本試劑混合物定位在次級試劑上。 5. Smaller chamber decompression causes the liquid sample to be further drawn along the channel, positioning the sample reagent mixture on the secondary reagent.

6.試劑再溶解及抗體-分析物-粒子標記物培育及結合時間。 6. Re-dissolution of the reagent and antibody-analyte-particle marker incubation and binding time.

7.第二次較小腔室減壓導致樣本進一步沿著通道移動，將樣本試劑混合物定位在第三試劑上 7. The second smaller chamber decompression causes the sample to move further along the channel, positioning the sample reagent mixture on the third reagent.

8.試劑再溶解及抗體-分析物-粒子標記物-順磁粒子培育及結合時間。 8. Re-dissolution of the reagent and antibody-analyte-particle marker-paramagnetic particle incubation and binding time.

9.將磁場施加至盒，定域至光學偵測區，將順磁粒子積聚至此位置，在每個通道中形成抗體-分析物-粒子標記物-順磁粒子複合物條帶。 9. Apply a magnetic field to the cartridge, localize to the optical detection zone, accumulate paramagnetic particles to this location, and form an antibody-analyte-particle marker-paramagnetic particle composite strip in each channel.

10.藉由腔室之再壓縮將樣本及未結合標記物從光學偵測區排出來將樣本液體及未結合標記物從偵測區移除 10. The sample liquid and the unbound label are removed from the optical detection zone by recompression of the chamber to remove the sample liquid and the unbound label from the detection zone.

11.讀數器之光學頭在整個帶材上掃描並且量測每個通道之螢光強度。螢光強度與肌鈣蛋白分析物濃度成比例。 11. The optical head of the reader scans the entire strip and measures the intensity of the fluorescence of each channel. Fluorescence intensity is proportional to the concentration of troponin analyte.

肌鈣蛋白I(TnI)檢定-試劑在圖14中識別 Troponin I (TnI) assay - reagent identified in Figure 14

步驟1：此為被動毛細管填充。TnI檢定使用兩個捕獲抗體，其中之每一者用生物素基團加標籤。標記物抗體用異硫氰酸螢光素(FITC)基團來加標籤。生物素基團及FITC分子充當第二及第三步驟之免疫原性標籤。 Step 1: This is a passive capillary fill. The TnI assay uses two capture antibodies, each of which is labeled with a biotin group. The marker antibody is labeled with a fluorescein isothiocyanate (FITC) group. The biotin group and the FITC molecule serve as immunogenic labels for the second and third steps.

步驟2：來自步驟一之樣本藉由流體讀數器控制(腔室減壓)來移動至次級試劑置放區域。此置放含有抗-FITC抗體塗布膠乳粒子。抗-FITC膠乳粒子結合FTIC標籤抗體(其結合至TnI複合物)。此反應係快速的。 Step 2: The sample from step 1 is moved to the secondary reagent placement area by fluid reader control (chamber decompression). This was placed containing anti-FITC antibody-coated latex particles. The anti-FITC latex particles bind to a FTIC tag antibody (which binds to the TnI complex). This reaction is fast.

步驟3：樣本藉由流體讀數器控制來移動至第三置放區。第三置放區域含有鏈黴親和素塗布磁性粒子。鏈黴親和素順磁粒子快速結合 生物素標記抗體，該等抗體結合至TnI複合物。在順磁粒子積聚之後，進行樣本/未結合標記物移除。然後執行螢光光學掃描。螢光強度與TnI濃度成比例。 Step 3: The sample is moved to the third placement zone by fluid reader control. The third placement area contains streptavidin coated magnetic particles. Streptavidin paramagnetic particles rapidly bind to biotinylated antibodies that bind to the TnI complex. After the paramagnetic particles have accumulated, sample/unbound label removal is performed. A fluorescent optical scan is then performed. The fluorescence intensity is proportional to the TnI concentration.

上述方法之示意圖在圖15示出。 A schematic diagram of the above method is shown in FIG.

步驟1係毛細管填充，步驟2及3在讀數器流體控制下。 Step 1 is capillary filling and steps 2 and 3 are under reader fluid control.

此方法係很有吸引力的，因為其具有通用應用並且極大地簡化檢定試劑，並且非常重要的是產生極好檢定效能(參見圖16(a)及(b)示出之示範性結果)，本發明方法在廣泛濃度範圍內係靈敏的。舉例而言，抗-FITC膠乳係用於其他檢定(例如，BNP)之通用標記物，同樣地鏈黴親和素順磁粒子在檢定之間亦係通用的。對於挑戰性檢定諸如TnI，試劑之分批生產變得容易得多。 This method is very attractive because it has a versatile application and greatly simplifies the assay reagents, and it is very important to produce excellent assay performance (see the exemplary results shown in Figures 16(a) and (b)), The method of the invention is sensitive over a wide range of concentrations. For example, anti-FITC latex is a universal marker for other assays (eg, BNP), and streptavidin paramagnetic particles are also commonly used between assays. For challenging assays such as TnI, batch production of reagents is much easier.

為了展示在空氣中執行光學偵測，諸如螢光偵測之有效性，發明人執行進一步C-肽檢定以展示如與空氣相比，在緩衝液或全血中進行時之響應。圖17展示在移除空氣之前(白色圓圈)及之後(黑色三角形)，使用本發明系統在緩衝液中之C-肽檢定響應。可以看出在不藉由空氣來移除液體的情況下，存在由於未結合標記物仍然存在於偵測區域中所導致的高背景。此在較低分析物濃度下導致不良精確度及靈敏度。在藉由空氣來移除液體之後，此未結合標記物得以有效地移除，留下很低背景，允許進行高靈敏度的量測。圖18展示在藉由空氣來移除液體之前(黑色圓圈)及之後(白色三角形)，使用本發明系統在全血中之C-肽檢定響應。可以看出在不藉由空氣來移除液體的情況下，存在由於未結合標記物仍然存在於偵測區域中所導致的高背景，並且沒有可見斜率，此歸因於使螢光量測淬滅的血液樣 本之干擾。在藉由空氣來移除液體之後，此未結合標記物及全血樣本有效地移除，在沒有干擾血細胞或未結合標記物之基本上無液體環境中留下結合試劑。此產生很低背景並且允許進行高靈敏度的量測。 To demonstrate the effectiveness of performing optical detection in air, such as fluorescence detection, the inventors performed a further C-peptide assay to demonstrate the response when performed in buffer or whole blood as compared to air. Figure 17 shows the C-peptide assay response in buffer prior to removal of air (white circles) and after (black triangles) using the system of the invention. It can be seen that in the case where the liquid is not removed by air, there is a high background due to the unbound label still being present in the detection area. This results in poor accuracy and sensitivity at lower analyte concentrations. This unbound label is effectively removed after removal of the liquid by air, leaving a very low background allowing for highly sensitive measurements. Figure 18 shows the C-peptide assay response in whole blood using the system of the invention before (black circles) and after (white triangles) removal of liquid by air. It can be seen that in the case where the liquid is not removed by air, there is a high background due to the unbound label still being present in the detection area, and there is no visible slope due to the quenching of the fluorescence amount. Interference with the extinguished blood sample. After removal of the liquid by air, the unbound label and the whole blood sample are effectively removed leaving a binding reagent in a substantially liquid-free environment that does not interfere with blood cells or unbound labels. This produces a very low background and allows for highly sensitive measurements.

本發明盒亦可進行不需要氣囊來進行檢定的檢定，例如，在判定血液樣本之凝血酶原時間(PT)及國際標準化比率(INR)中。PT及INR係評估凝血之外因性途徑(PT/INR)的檢定。其用於在量測華法林劑量、肝損傷及維生素K狀態中判定血液之凝血趨勢。 The kit of the present invention can also perform assays that do not require an air bag for verification, for example, in determining prothrombin time (PT) and international normalization ratio (INR) of a blood sample. PT and INR were used to assess the coagulation-induced pathogen (PT/INR). It is used to determine the blood coagulation tendency in measuring warfarin dose, liver damage and vitamin K status.

圖19示出PT/INR量測之方法比較繪圖，其使用不具有藉由氣體腔室提供之任何流體控制的通道來產生。在此方面，樣本僅藉由毛細管作用來填充。為免生疑問，PT/INR量測亦可使用具有相關聯氣體腔室之通道來進行，允許樣本之流體控制，從而產生標準化填充速率。與先前描述免疫檢定實例相比，在帶材之偵測區域中，通道得以加寬以便允許增加將要檢定之樣本之體積。另外，INR/PT特異性試劑置放在此區域中。試劑含有啟始外因性凝血級聯之所有所需組分及特異性凝血酶螢光團受質，其藉由凝血酶從非螢光形式轉換成螢光形式。毛細管填充將試劑再懸浮並且允許偵測凝血酶活性。所量測的凝血酶活性(螢光強度)用於判定PT/INR結果。 Figure 19 shows a comparative plot of method for PT/INR measurements that is generated using channels that do not have any fluid control provided by the gas chamber. In this respect, the sample is only filled by capillary action. For the avoidance of doubt, PT/INR measurements can also be made using channels with associated gas chambers, allowing fluid control of the sample to produce a standardized fill rate. In the detection zone of the strip, the channel is widened to allow for an increase in the volume of the sample to be assayed, as compared to the previously described immunoassay example. In addition, INR/PT specific reagents are placed in this region. The reagent contains all of the desired components of the initiation of the exogenous coagulation cascade and a specific thrombin fluorophore receptor that is converted from a non-fluorescent form to a fluorescent form by thrombin. Capillary filling resuspends the reagent and allows detection of thrombin activity. The measured thrombin activity (fluorescence intensity) was used to determine the PT/INR results.

Claims (65)

一種用於對於一液體樣本進行一檢定的自給式微流控系統，該微流控系統包含：用於接收該液體樣本之一樣本輸入埠，該樣本輸入埠連接至至少一個微流控通道，其中每個/該等微流控通道包含置放在其中的用於進行該檢定的一或多個試劑及一偵測區，該偵測區用於偵測可存在於一樣本中之任何分析物或其分析物反應產物；及每個/該等微流控通道進一步流體連接至在每個/該偵測區下游的一可壓縮、氣體填充腔室，及其中該系統由三個層形成，該等層層疊在一起以界定每個/該等微流控通道及該氣體填充腔室，並且其中使該腔室壓縮或減壓導致將氣體從該腔室中排出或抽吸至該腔室中，進而導致該/每個微流控通道內之該液體樣本之運動。  A self-contained microfluidic system for performing an assay on a liquid sample, the microfluidic system comprising: a sample input port for receiving the liquid sample, the sample input port being coupled to at least one microfluidic channel, wherein Each/the microfluidic channel includes one or more reagents and a detection zone disposed therein for performing the assay, the detection zone for detecting any analyte that may be present in the same Or an analyte reaction product thereof; and each/the microfluidic channel is further fluidly coupled to a compressible, gas-filled chamber downstream of each/the detection zone, and wherein the system is formed of three layers, The layers are stacked together to define each/the microfluidic channel and the gas-filled chamber, and wherein compressing or decompressing the chamber results in venting or pumping gas from the chamber to the chamber Medium, which in turn causes movement of the liquid sample within the/each microfluidic channel.   如申請專利範圍第1項之微流控系統，其中在該液體樣本與置放在該/每個微流控通道內之該一或多個試劑反應之後，從該腔室排出之氣體用來將液體從該/每個微流控通道內之該偵測區中移除，以使得可在一實質上無液體環境中偵測該/每個偵測區內之任何分析物或分析物反應產物。  The microfluidic system of claim 1, wherein the gas discharged from the chamber is used after the liquid sample reacts with the one or more reagents placed in the/each microfluidic channel Removing liquid from the detection zone within the/each microfluidic channel such that any analyte or analyte reaction within the/detection zone can be detected in a substantially liquid-free environment product.   如申請專利範圍第1或2項之微流控系統，其包含複數個微流控通道，其中該等複數個微流控通道中之每一者與該樣本輸入埠流體連通，視需要其中一單一微流控通道劃分成該等複數個通道。  The microfluidic system of claim 1 or 2, comprising a plurality of microfluidic channels, wherein each of the plurality of microfluidic channels is in fluid communication with the sample input port, one of the A single microfluidic channel is divided into the plurality of channels.   如申請專利範圍第3項之微流控系統，其中該等複數個微流控通道中之每一者連接至一相應氣體填充腔室，且/或兩個或兩個以上微流控通道 連接至一氣體填充腔室。  The microfluidic system of claim 3, wherein each of the plurality of microfluidic channels is connected to a corresponding gas-filled chamber, and/or two or more microfluidic channels are connected To a gas filled chamber.   如申請專利範圍第1-4項中任一項之微流控系統，其中該樣本埠連接至該等/每個微流控通道之一第一端並且該等/每個微流控通道之一第二端連接至該等氣體填充腔室中之一或多者。  The microfluidic system of any one of claims 1-4, wherein the sample is coupled to one of the first ends of the/each microfluidic channel and the/each microfluidic channel A second end is coupled to one or more of the gas filled chambers.   如申請專利範圍第1-5項中任一項之微流控系統，其進一步包含被設計來接收流體廢物及/或過量液體樣本的一或多個吸收器特徵。  The microfluidic system of any of claims 1-5, further comprising one or more absorber features designed to receive fluid waste and/or excess liquid sample.   如申請專利範圍第1-6項中任一項之微流控系統，其中該等頂部及底部層係平面的並且視需要具有均勻厚度。  The microfluidic system of any one of claims 1-6, wherein the top and bottom layers are planar and optionally have a uniform thickness.   如申請專利範圍第7項之微流控系統，其中該等頂部及底部層由相同材料形成。  The microfluidic system of claim 7, wherein the top and bottom layers are formed of the same material.   如申請專利範圍第7或8項之微流控系統，其中該系統由一捲筒或卷對卷製程來形成。  A microfluidic system as claimed in claim 7 or 8, wherein the system is formed by a reel or roll-to-roll process.   如申請專利範圍第7-9項中任一項之微流控系統，其中該等平面基板藉由施加熱及/或使用黏著劑來密封在一起。  The microfluidic system of any one of claims 7-9, wherein the planar substrates are sealed together by applying heat and/or using an adhesive.   如申請專利範圍第10項之微流控系統，其中該等平面基板使用一黏著劑來密封在一起，該黏著劑有彈性並且促進每個/該腔室之可壓縮性。  A microfluidic system according to claim 10, wherein the planar substrates are sealed together using an adhesive which is elastic and promotes compressibility of each of the chambers.   如申請專利範圍第1-11項中任一項之微流控系統，其中該系統中之該等/每個微流控通道包含一或多個流體止擋特徵，其被設計來防止該樣本及/或其他流體僅藉助於毛細管作用來經過該止擋特徵。  The microfluidic system of any of claims 1-11, wherein the/each microfluidic channel in the system comprises one or more fluid stop features designed to prevent the sample And/or other fluids pass through the stop feature only by capillary action.   如申請專利範圍第1-12項中任一項之微流控系統，其包含一單向閥，該單向閥被設計來僅在藉由毛細管作用將一液體樣本引入該系統中時允許氣體退出該系統，同時不允許經由該閥將流體引入該系統中。  A microfluidic system according to any one of claims 1 to 12, which comprises a one-way valve designed to allow gas only when a liquid sample is introduced into the system by capillary action The system is withdrawn while not allowing fluid to be introduced into the system via the valve.   如申請專利範圍第13項之微流控系統，其中該閥相鄰於一止擋特徵來定位，該止擋特徵被設計來防止該樣本僅藉由毛細管作用在該微流控通道內進一步傳送。  The microfluidic system of claim 13, wherein the valve is positioned adjacent to a stop feature, the stop feature being designed to prevent the sample from being further transferred within the microfluidic channel only by capillary action .   如申請專利範圍第14項之微流控系統，其中該閥定位在具有比該/每個微流控通道更小之尺寸的一微流控通道中並且與該等微流控通道中之一者流體連通。  The microfluidic system of claim 14, wherein the valve is positioned in a microfluidic channel having a smaller size than the/each microfluidic channel and is associated with one of the microfluidic channels The fluid is connected.   如申請專利範圍第1-15項中任一項之微流控系統，其包含與該等/每個通道接觸之一或多個電極特徵，其用於量測或偵測存在於該等/每個通道中之液體。  The microfluidic system of any of claims 1-15, comprising one or more electrode features in contact with the/each channel for measuring or detecting presence in the / The liquid in each channel.   如申請專利範圍第1-16項中任一項之微流控系統，其進一步包含置放在該通道內之一分析物結合劑，其中視需要該分析物結合劑結合至該通道之一表面。  The microfluidic system of any one of claims 1 to 16, further comprising an analyte binding agent disposed in the channel, wherein the analyte binding agent is bound to one surface of the channel as needed .   如申請專利範圍第17項之微流控系統，其中該結合劑附接至一磁性或順磁粒子。  The microfluidic system of claim 17, wherein the binder is attached to a magnetic or paramagnetic particle.   如申請專利範圍第17或18項之微流控系統，其中該結合劑或磁性/順磁粒子置放在該系統之該等/每個微流控通道內，以使得在該樣本施加至該系統並且被抽吸至該等/每個通道中之後，該等黏合劑或磁性/順磁粒子藉由該液體樣本來懸浮。  The microfluidic system of claim 17 or 18, wherein the binder or magnetic/paramagnetic particles are placed in the/each microfluidic channel of the system such that the sample is applied to the sample After the system is pumped into the/each channel, the binder or magnetic/paramagnetic particles are suspended by the liquid sample.   如申請專利範圍第17-19項中任一項之微流控系統，其中該結合劑或磁性/順磁粒子置放在該等/每個微流控通道之一區域內，該區域藉由在該置放區域之任一端處之特徵來界定，該等特徵被設計來在該等磁性/順磁粒子最初置放在該/每個通道中時限制該等粒子之運動。  The microfluidic system of any one of claims 17 to 19, wherein the binder or magnetic/paramagnetic particles are placed in a region of the/each microfluidic channel, the region The features are defined at either end of the placement area and are designed to limit the movement of the magnetic/paramagnetic particles as they are initially placed in the/each channel.   如申請專利範圍第19或20項之微流控系統，其中該等磁性/順磁粒子置放在與一磁鐵或磁力緊密鄰近之該系統之外表面相反的該/每個通道之一內表面上。  The microfluidic system of claim 19 or 20, wherein the magnetic/paramagnetic particles are placed on an inner surface of the/each channel opposite the surface of the system in close proximity to a magnet or magnetic force on.   如申請專利範圍第1-21項中任一項之微流控系統，其中該系統進一步包含置放在該等/每個微流控通道內之一或多個額外試劑，該等額外試劑促進存在於該樣本中之分析物之偵測。  The microfluidic system of any one of claims 1 to 21, wherein the system further comprises one or more additional reagents disposed in the/each microfluidic channel, the additional reagents being promoted Detection of analytes present in the sample.   如申請專利範圍第22項之微流控系統，其中該一或多個額外試劑包括一標記物，該標記物適於特異性結合至將要偵測之一分析物以促進分析物偵測。  A microfluidic system according to claim 22, wherein the one or more additional reagents comprise a label adapted to specifically bind to one of the analytes to be detected to facilitate analyte detection.   如申請專利範圍第22或23項之微流控系統，其中經由將氣體抽吸回到該/每個氣體填充腔室中，分析物在該等/每個微流控通道之一第一區域中結合至該分析物結合劑，然後被傳送至該等/每個微流控通道之另外一或多個區域，其中置放該一或多個其他試劑及/或標記物。  A microfluidic system according to claim 22 or 23, wherein the analyte is in the first region of one of the per microfluidic channels by pumping gas back into the/each gas-filled chamber The analyte binding agent is bound to the other or regions of the microfluidic channel, wherein the one or more additional reagents and/or labels are placed.   如申請專利範圍第1-24項中任一項之微流控系統，其中該系統能夠對於一單一樣本執行複數個(諸如2、3、4、5、6、7、8、9、10或更多個)相同及/或不同檢定。  The microfluidic system of any one of claims 1 to 24, wherein the system is capable of performing a plurality of samples (such as 2, 3, 4, 5, 6, 7, 8, 9, 10 or More) Same and/or different checks.   如申請專利範圍第1-25項中任一項之微流控系統，其中施加至該系統之該樣本之體積小於100 μl、50 μl，諸如小於40 μl、30 μl或20 μl。 The microfluidic system of any one of claims 1 to 25, wherein the sample applied to the system has a volume of less than 100 μl , 50 μl , such as less than 40 μl , 30 μl or 20 μ. l. 一種套組，其包含如申請專利範圍第1-26項中任一項之微流控系統，以及一樣本收集裝置。  A kit comprising the microfluidic system of any one of claims 1-26, and the same collection device.   如申請專利範圍第27項之套組，其中該樣本收集裝置適於***該系統之樣本輸入埠並且其後為該輸入埠提供一密封。  A kit of claim 27, wherein the sample collection device is adapted to be inserted into a sample input port of the system and thereafter provide a seal for the input port.   如申請專利範圍第28項之套組，其用於進行一核酸偵測檢定。  For example, the kit of claim 28 is used for performing a nucleic acid detection test.   一種與如申請專利範圍第1-26項中任一項之微流控系統，或如申請專利範圍第27-29項之套組一起使用的讀數器裝置，該讀數器裝置包含：力控制構件，其用於控制該微流控系統之一氣體填充腔室之壓縮或減壓；及偵測構件，其用於實現引入該微流控盒中之一液體樣本內之一所需分析物，或其分析物反應產物的偵測；其中該力控制構件包含一壓電彎曲致動器，其被設計來直接地或經由該致動器之位移來間接地實現該氣體填充腔室之壓縮或減壓。  A reader device for use with a microfluidic system according to any one of claims 1-26, or a kit as claimed in claims 27-27, the reader device comprising: a force control member And for detecting compression or decompression of a gas-filled chamber of the microfluidic system; and detecting means for implementing an analyte required to introduce one of the liquid samples in the microfluidic cartridge, Or detection of an analyte reaction product thereof; wherein the force control member comprises a piezoelectric bending actuator designed to indirectly effect compression of the gas-filled chamber, either directly or via displacement of the actuator stress reliever.   如申請專利範圍第30項之讀數器裝置，其中該壓電彎曲機呈一帶材、棒、桿等形式，其包含一第一固定端及一第二未固定端，其中在發出適當電信號後，該第二未固定端可自由彎曲遠離該氣體填充腔室。  The reader device of claim 30, wherein the piezoelectric bending machine is in the form of a strip, a rod, a rod, or the like, and includes a first fixed end and a second unfixed end, wherein after issuing an appropriate electrical signal The second unfixed end is free to bend away from the gas filled chamber.   如申請專利範圍第31項之讀數器裝置，其進一步包含一底座，該底座能夠與該氣體填充腔室之一外表面嚙合，其中該底座之一頂部表面與該壓電彎曲機接觸並且其中該底座能夠經由該彎曲機之作用來實現該氣體填充腔室之壓縮或減壓。  The reader device of claim 31, further comprising a base engageable with an outer surface of one of the gas-filled chambers, wherein a top surface of the base is in contact with the piezoelectric bending machine and wherein The base can effect compression or decompression of the gas-filled chamber via the action of the bending machine.   如申請專利範圍第30-32項中任一項之讀數器裝置，其中該壓電彎曲機最初偏置，或導致該相關聯底座與該氣體填充腔室之外表面接觸。  The reader device of any one of claims 30-32, wherein the piezoelectric bending machine is initially biased or causes the associated base to contact the outer surface of the gas-filled chamber.   如申請專利範圍第30-33項之讀數器裝置，其進一步包含偵測構件，其用於實現引入該微流控系統中之一液體樣本內之一所需分析物或其分析物反應產物的偵測。  A reader device according to claims 30-33, further comprising a detecting member for effecting introduction of an analyte or an analyte reaction product thereof in one of the liquid samples in the microfluidic system Detection.   如申請專利範圍第30-33項之讀數器裝置，其進一步包含一接收埠，該接收埠適於接收不同大小之系統，每個不同大小之系統被設計來執行規 定數目之檢定。  A reader device as claimed in claims 30-33, further comprising a receiving cassette adapted to receive systems of different sizes, each system of different size being designed to perform a specified number of checks.   如申請專利範圍第35項之讀數器裝置，其中該接收埠經如此調適以確保每個不同大小之系統的正確***及識別。  The reader device of claim 35, wherein the receiving device is adapted to ensure proper insertion and identification of each of the different sized systems.   如申請專利範圍第30-36項之讀數器裝置，其進一步包含緊密鄰近被引入該讀數器中之如申請專利範圍第18-26項之系統的一永磁鐵或被設計來向該系統施加一磁場的電磁鐵，以便該等磁性/順磁粒子集中並保持在該系統之該/每個微流控通道之該偵測區中。  A reader device as claimed in claim 30-36, further comprising a permanent magnet in close proximity to the system incorporated in the reader as claimed in claims 18-26 or designed to apply a magnetic field to the system Electromagnets such that the magnetic/paramagnetic particles are concentrated and held in the detection zone of the/each microfluidic channel of the system.   如申請專利範圍第30-37中任一項之讀數器裝置，其中該壓電彎曲機或底座被設計來僅接觸氣體填充腔室之該總外表面之一部分。  The reader device of any of claims 30-37, wherein the piezoelectric bender or base is designed to contact only a portion of the total outer surface of the gas-filled chamber.   如申請專利範圍第38項之讀數器裝置，其中每個壓電彎曲機或底座經設定大小來接觸每個腔室之10與50%之間之外表面。  A reader device according to claim 38, wherein each piezoelectric bending machine or base is sized to contact between 10 and 50% of the outer surface of each chamber.   如申請專利範圍第30-39項中任一項之讀數器裝置，其中該壓電彎曲機被設計來使用電路來彎曲及鬆弛，該電路存在於該讀數器中並且連接至該壓電彎曲機。  The reader device of any one of claims 30-39, wherein the piezoelectric bending machine is designed to bend and relax using an electrical circuit, the circuit being present in the reader and coupled to the piezoelectric bending machine .   如申請專利範圍第40項之讀數器裝置，其中該電路能夠導致該壓電彎曲機以一可變速率來彎曲以使得該系統內之氣體可以不同速率被抽吸至該/每個氣體填充腔室中及/或從其中排出。  The reader device of claim 40, wherein the circuit is capable of causing the piezoelectric bending machine to bend at a variable rate such that gas within the system can be drawn to the/each gas filled chamber at different rates Discharged from and/or from the chamber.   如申請專利範圍第30-41項之讀數器裝置，其中該偵測構件係一光學偵測裝置，諸如一螢光計或分光光度計。  The reader device of claim 30-41, wherein the detecting member is an optical detecting device such as a fluorometer or a spectrophotometer.   如申請專利範圍第30-43項中任一項之讀數器裝置，其進一步包含允許在一特定溫度，或複數個溫度下進行檢定之加熱及/或冷卻構件。  The reader device of any one of claims 30-43, further comprising a heating and/or cooling member that allows for verification at a particular temperature, or a plurality of temperatures.   一種包含一自給式微流控系統及一相關聯讀數器裝置的檢定系 統，其中：該自給式微流控系統包含：一樣本輸入埠，其用於接收將要檢定之一液體樣本，該樣本輸入埠連接至至少一個微流控通道，其中每個/該等微流控通道包含置放在其中以用於進行一檢定的一或多個試劑及一偵測區，該偵測區用於偵測可存在於一樣本中之任何分析物或分析物反應產物；及每個/該等微流控通道與每個/該偵測區下游之一可壓縮、氣體填充腔室流體連通，其中該微流控系統由三個層形成，該等層層疊在一起以界定每個/該等微流控通道及該氣體填充腔室，並且其中使該腔室壓縮或減壓導致將氣體從該腔室中排出或抽吸至該腔室中，進而導致該/每個微流控通道內之該液體樣本之運動；及與該微流控系統一起使用之一讀數器裝置，該讀數器裝置包含：力控制構件，其用於控制該微流控系統之氣體填充腔室之壓縮或減壓；及偵測構件，其用於實現引入該微流控盒中之一液體樣本內之一所需分析物，或其分析物反應產物的偵測；其中該力控制構件包含一壓電彎曲致動器，其被設計來直接地或經由該致動器之位移來間接地將該氣體填充腔室壓縮或減壓。  An assay system comprising a self-contained microfluidic system and an associated reader device, wherein: the self-contained microfluidic system comprises: the same input port for receiving a liquid sample to be assayed, the sample input port connection Up to at least one microfluidic channel, wherein each/the microfluidic channel includes one or more reagents and a detection zone disposed therein for performing an assay, the detection zone being used for detecting Any analyte or analyte reaction product present in the same; and each/the microfluidic channel is in fluid communication with a compressible, gas-filled chamber downstream of each/the detection zone, wherein the microfluid The control system is formed of three layers that are stacked together to define each/the microfluidic channel and the gas-filled chamber, and wherein compressing or decompressing the chamber results in gas being drawn from the chamber Discharging or pumping into the chamber, thereby causing movement of the liquid sample within the/each microfluidic channel; and using a reader device with the microfluidic system, the reader device comprising: a force Control member for Compressing or decompressing a gas-filled chamber of the microfluidic system; and detecting means for effecting introduction of one of the analytes in one of the liquid samples in the microfluidic cartridge, or an analyte reaction thereof Detection of the product; wherein the force control member comprises a piezoelectric bending actuator designed to indirectly compress or decompress the gas filled chamber directly or via displacement of the actuator.   一種對於一液體樣本進行一檢定之方法，該方法包含：a)將如本文描述之一微流控系統提供至如本文描述之一讀數器裝置；b)壓縮該微流控系統之一/該等氣體填充腔室，以便將氣體從該等/每個氣體填充腔室排出； c)將一液體樣本引入該微流控系統並且允許藉由毛細管作用將該樣本抽吸至該等/每個微流控通道中，且/或部分地將該等/每個氣體填充腔室減壓以使得氣體被抽吸至該等/每個腔室中，從而導致該液體樣本被抽吸至該等/每個微流控通道中；d)允許一或多個試劑與存在於該液體樣本中之任何分析物反應；e)視需要部分地進一步部分地將該微流控系統之該等/每個氣體填充腔室減壓，以使得該液體樣本進一步沿著該等/每個微流控通道朝向該等/每個氣體填充腔室被抽吸並且視需要使該液體樣本與一分析物結合劑及/或一或多個其他試劑接觸；f)視需要捕獲任何分析物或分析物反應產物並且壓縮該等/每個氣體填充腔室，以使得從該等/每個腔室排出之氣體導致該液體樣本及未捕獲材料被推動遠離任何所捕獲分析物或分析物反應產物；及g)偵測任何分析物或分析物反應產物，或所捕獲的分析物或分析物反應產物。  A method for performing a assay on a liquid sample, the method comprising: a) providing a microfluidic system as described herein to one of the reader devices as described herein; b) compressing one of the microfluidic systems/the The gas fills the chamber to expel gas from the/each gas-filled chamber; c) introduces a liquid sample into the microfluidic system and allows the sample to be drawn to the/each by capillary action In the microfluidic channel, and/or partially decompressing the gas/each gas filling chamber such that gas is drawn into the/each chamber, causing the liquid sample to be drawn to the / in each microfluidic channel; d) allowing one or more reagents to react with any analyte present in the liquid sample; e) partially partially further/pertheming the microfluidic system The gas-filled chamber is depressurized such that the liquid sample is further drawn along the/each microfluidic channel toward the/each gas-filled chamber and the liquid sample is combined with an analyte as needed And/or one or more other reagents are contacted; f) as needed Obtaining any analyte or analyte reaction product and compressing the/each gas-filled chamber such that gas exiting the chamber/each chamber causes the liquid sample and the uncaptured material to be pushed away from any captured analyte Or an analyte reaction product; and g) detecting any analyte or analyte reaction product, or the captured analyte or analyte reaction product.   如申請專利範圍第45項之方法，其中步驟e)作為單一或多個步驟來執行，其中對應於力之減少的執行次數，該樣本相應地被抽吸至該/每個微流控通道內之另一個或多個連續位置。  The method of claim 45, wherein step e) is performed as a single or multiple steps, wherein the sample is correspondingly drawn into the/each microfluidic channel corresponding to the number of executions of the reduced force Another or more consecutive locations.   如申請專利範圍第45或46項之方法，其中所形成的該分析物/分析物結合劑複合物或分析物反應產物/分析物結合劑複合物包含磁性或順磁粒子。  The method of claim 45 or 46, wherein the analyte/analyte binder complex or analyte reaction product/analyte binder complex formed comprises magnetic or paramagnetic particles.   如申請專利範圍第47項之方法，其中用於形成該等複合物之該等磁性粒子最初置放在與一磁鐵緊密接觸，或施加一磁力的該系統之外表面相 反的該(該等)微流控通道之一內表面上，以使得該等磁性粒子被側向地抽吸穿過該(該等)微流控通道。  The method of claim 47, wherein the magnetic particles used to form the composite are initially placed in intimate contact with a magnet or a magnetic force is applied to the outer surface of the system. One of the microfluidic channels is on the inner surface such that the magnetic particles are drawn laterally through the (micro)fluidic channel.   如申請專利範圍第45-48項之方法，其中步驟e)作為單一或多個步驟來執行，其中對應於力之減少的執行次數，該樣本相應地被抽吸至該/每個微流控通道內之另一個或多個連續位置。  The method of claim 45, wherein step e) is performed as a single or multiple steps, wherein the sample is correspondingly pumped to the/each microfluidic corresponding to the number of executions of the reduced force Another or more consecutive locations within the channel.   如申請專利範圍第45-49項中任一項之方法，其中導致從捕獲該分析物/分析物結合劑複合物之該(該等)微流控通道之至少一部分排出液體的從該(該等)腔室排出的氣體之體積足以導致該液體從該偵測區或其一部分移除，但是不進一步沿著該(該等)微流控通道移除。  The method of any one of claims 45-49, wherein the liquid is discharged from at least a portion of the microfluidic channel that captures the analyte/analyte binder complex (from the The volume of gas exiting the chamber is sufficient to cause the liquid to be removed from the detection zone or a portion thereof, but not further along the microfluidic channel.   一種用於進行複數個不同檢定的自給式拋棄式微流控系統，該微流控盒包含：用於將一液體樣本引入該微流控盒中之一樣本輸入埠；多個微流控通道；該等微流控通道中之每一者適於接收該液體樣本之一部分並且能夠使用一或多個試劑來對於樣本之該部分進行一或多個檢定，該等試劑在引入液體樣本之前存在於該等微流控通道中之每一者內；及其中每個微流控通道內之流體運動藉由該微流控系統之兩個或兩個以上氣體填充腔室之壓縮及/或減壓來獨立地可控制，該等腔室各自與該等微流控通道中之一或多者流體連通。  A self-contained disposable microfluidic system for performing a plurality of different assays, the microfluidic box comprising: a sample input port for introducing a liquid sample into the microfluidic box; and a plurality of microfluidic channels; Each of the microfluidic channels is adapted to receive a portion of the liquid sample and can use one or more reagents to perform one or more assays on the portion of the sample, the reagents being present prior to introduction of the liquid sample Each of the microfluidic channels; and fluid movement within each of the microfluidic channels; compression and/or decompression of the chamber by two or more gases of the microfluidic system Independently controllable, the chambers are each in fluid communication with one or more of the microfluidic channels.   如申請專利範圍第51項之自給式拋棄式微流控系統，其用於如申請專利範圍第45-50項中任一項之方法中。  The self-contained disposable microfluidic system of claim 51, which is used in the method of any one of claims 45-50.   如申請專利範圍第51項之自給式拋棄式微流控系統，其進一步包含 如申請專利範圍第1-26項所定義之特徵。  The self-contained disposable microfluidic system of claim 51, further comprising the features as defined in claim 1-26.   如申請專利範圍第51-53項中任一項之自給式拋棄式微流控系統，其能夠執行以下類型之檢定中之至少兩者、三者、四者、五者或更多者：免疫檢定、核酸檢定、基於受體之檢定、細胞計數檢定、比色檢定、酶檢定、電泳檢定、電化學檢定、光譜檢定、層析檢定、顯微鏡檢定、局部檢定(topographic assay)、量熱檢定、濁度檢定、凝集檢定、黏度檢定、凝血檢定、凝結時間檢定、蛋白質合成檢定、組織學檢定、培養檢定或滲透壓檢定。  A self-contained disposable microfluidic system according to any one of claims 51-53, which is capable of performing at least two, three, four, five or more of the following types of assays: immunoassay , nucleic acid assay, receptor-based assay, cell count assay, colorimetric assay, enzyme assay, electrophoretic assay, electrochemical assay, spectral assay, chromatographic assay, microscopic assay, topographic assay, calorimetric assay, turbidity Degree test, agglutination test, viscosity test, coagulation test, coagulation time test, protein synthesis test, histological test, culture test or osmotic pressure test.   如申請專利範圍第50-53項中任一項之自給式拋棄式微流控系統，其能夠進行一組單獨檢定，該等檢定被設計來測試一心臟病狀、妊娠、腎病狀、神經學病狀、一腎上腺病狀、一肝臟病狀、糖尿病、病原體或藥物濫用。  A self-contained disposable microfluidic system, as claimed in any one of claims 50-53, capable of performing a separate set of tests designed to test a heart condition, pregnancy, kidney condition, neurological disease Symptoms, an adrenal gland condition, a liver condition, diabetes, pathogens or drug abuse.   如申請專利範圍第55項之自給式拋棄式微流控系統，其用於偵測一心臟病狀並且其中該組單獨檢定用於偵測以下中之一或多者：脂質水準、脂蛋白元；升半胱胺酸；C-反應蛋白(CRP)肌鈣蛋白，BNP；及/或心臟酶。  A self-contained disposable microfluidic system as claimed in claim 55, for detecting a heart condition and wherein the set of individual assays is for detecting one or more of the following: lipid level, lipoprotein; C-cysteine; C-reactive protein (CRP) troponin, BNP; and/or cardiac enzyme.   如申請專利範圍第55項之自給式拋棄式微流控系統，其用於偵測一腎上腺病狀，並且其中該組單獨檢定用於偵測醛固酮、皮質醇、18-羥皮質醇及/或DHEA-S中之一或多者。  A self-contained disposable microfluidic system as claimed in claim 55, for detecting an adrenal gland condition, and wherein the set is individually assayed for detecting aldosterone, cortisol, 18-hydroxycortisol and/or DHEA One or more of -S.   如申請專利範圍第55項之自給式拋棄式微流控系統，其用於偵測一肝臟病狀並且其中該組單獨檢定用於偵測一或多種肝臟酶、膽紅素、白蛋白、凝血酶原之一水準及/或一或多種病毒之存在。  A self-contained disposable microfluidic system as claimed in claim 55, for detecting a liver condition and wherein the set of individual assays is for detecting one or more liver enzymes, bilirubin, albumin, thrombin One of the original levels and / or the presence of one or more viruses.   如申請專利範圍第55項之自給式拋棄式微流控系統，其用於偵測處 於患上糖尿病之風險中之受試者或確認患有糖尿病之受試者並且其中該組單獨檢定用於偵測以下中之一或多者：脂質水準、全血計數、禁食葡萄糖水準、血紅蛋白A1c及/或白蛋白。  A self-contained disposable microfluidic system as claimed in claim 55, which is for detecting a subject at risk of developing diabetes or a subject confirmed to have diabetes and wherein the group is individually tested for detection One or more of the following are measured: lipid level, whole blood count, fasting glucose level, hemoglobin A1c and/or albumin.   如申請專利範圍第55項之自給式拋棄式微流控系統，其用於偵測藥物濫用，其中該組檢定用於偵測以下中之一或多者：***；巴比妥酸鹽；丁基原啡因；苯二氮平；古柯鹼；搖頭丸；甲基***；***(鴉片劑/嗎啡)；***；三環抗抑鬱藥；及/或***。  For example, the self-contained disposable microfluidic system of claim 55 is used for detecting drug abuse, wherein the group of tests is for detecting one or more of the following: amphetamine; barbiturate; butyl morphine Because of; benzodiazepine; ***e; ecstasy; methamphetamine; heroin (opiates / morphine); methadone; tricyclic antidepressants; and / or cannabis.   一種用於進行多組檢定的多重檢定平臺，該多重檢定平臺包含如申請專利範圍第50-60項中任一項之複數個微流控系統，每個系統能夠對於一樣本進行一組規定檢定及被構建來能夠接收及檢驗該等複數個微流控系統中之每一者的一讀數器，其中該讀數器可被組配來偵測及/或判定可存在於該樣本中之一組分析物之水準。  A multi-validation platform for performing a plurality of sets of assays, the multi-assay platform comprising a plurality of microfluidic systems as claimed in any one of claims 50-60, each system being capable of performing a set of prescribed tests on the same And a reader configured to receive and verify each of the plurality of microfluidic systems, wherein the reader can be configured to detect and/or determine a group that can be present in the sample The level of analytes.   如申請專利範圍第61項之用於進行多組檢定之多重檢定平臺，其與如申請專利範圍第30-43項中任一項之讀數器裝置一起使用。  A multi-assay platform for performing a plurality of sets of assays, as claimed in claim 61, for use with a reader device according to any one of claims 30-43.   一種根據如申請專利範圍第1-26項中任一項之檢定系統來使用的閥系統，該閥系統包含：在如本發明之檢定系統之一頂部或底部表面中之一通氣孔或縫隙開口；及通向該檢定系統之該微流控通道的具有較小尺寸之一微流控通道，該較小尺寸之微流控通道與該檢定系統之該通氣孔或縫隙開口及該微流控通道流體連通。  A valve system for use in an assay system according to any one of claims 1 to 26, the valve system comprising: a vent or slit opening in a top or bottom surface of one of the assay systems of the present invention; And a microfluidic channel having a smaller size to the microfluidic channel of the assay system, the smaller size microfluidic channel and the vent or slit opening of the assay system and the microfluidic channel Fluid communication.   如申請專利範圍第63項之閥系統，其中該閥系統被定位成與該微流 控通道之一毛細管止擋物相鄰，以使得在該樣本引入該檢定系統後，該樣本藉由毛細管作用來填充直至該毛細管止擋物並且該樣本之一部分亦至少部分地填充該較小尺寸之微通道。  A valve system according to claim 63, wherein the valve system is positioned adjacent to a capillary stop of the microfluidic channel such that after the sample is introduced into the assay system, the sample is acted upon by capillary action To fill the capillary stop and a portion of the sample also at least partially fills the smaller sized microchannel.   如申請專利範圍第64項之閥系統，其中至少部分地填充該較小尺寸之微通道的該樣本之部分充當沿著該較小尺寸之微通道之進一步流體傳送及經由該通氣孔或縫隙之流體輸出的屏障。  The valve system of claim 64, wherein the portion of the sample at least partially filling the smaller sized microchannel acts as a further fluid transfer along the smaller sized microchannel and through the vent or slit A barrier to fluid output.