WO2015198097A1

WO2015198097A1 - System and method for analysis of analytes in samples

Info

Publication number: WO2015198097A1
Application number: PCT/IB2014/062621
Authority: WO
Inventors: Vijaywanth MATHUR; Vaishali NEVREKER; Bikram Bhowmik; Dhanada DESHPANDE; Jignesh SAVANI; Prakash BAPODARA; Rohit Gupta
Original assignee: Diasys Diagnostics India Private Limited
Priority date: 2014-06-26
Filing date: 2014-06-26
Publication date: 2015-12-30

Abstract

The present invention relates to a system and a method for testing of disease markers for diseases such as diabetic, cardiac, thyroid and infectious diseases, in biological samples using lab-on-cartridge based electrochemical ELISA platform technology, which is common to a typical large laboratory instruments for rapid quantitative measurement that can expedite treatment of patients and contribute to improved outcomes, including enhanced physician and patient satisfaction.

Description

SYSTEM AND METHOD FOR ANALYSIS OF ANALYTES IN SAMPLES

TECHNICAL FIELD

The present invention relates to a system and a method for rapid quantitative measurement of disease markers in biological samples. Particularly, the present invention provides a handheld, point-of-care device for the measurement of clinical diagnostic parameters using lab-on- cartridge based electrochemical ELISA (biosensor/immunosensor) platform technology.

BACKGROUND Background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication, specifically or implicitly referenced is prior art.

The identification of a disease or medical condition of a patient using various diagnostic tests plays an important role in the treatment of the patients. The doctors typically validate diagnostic test results rather than starting treatment for a disease based on clinical symptoms. Therefore, in most cases, timely availability of diagnostic test results is crucial, not only for well- being of the patient, but also for achieving an efficient overall cost of the treatment.

Most of the diagnostic tests related to blood or other biological fluids are based on immunoassay, which is a biochemical test that measures presence of and/or concentration of a macromolecule, often referred to as an analyte, in a solution through the use of an antibody or immunoglobulin. Immunoassays rely on the ability of an antibody to recognize and bind a specific macromolecule.

In addition to the binding of an antibody to its antigen, the other key feature of all immunoassays is a means to produce a measurable signal in response to the binding. Most, though not all, immunoassays involve chemically linking antibodies or antigens with some kind of detectable labels. A large number of labels exist in modern immunoassays and they allow for detection through different means. Possibly one of the most popular labels that are used in immunoassays is enzymes. The immunoassays which employ enzymes are referred to as Enzyme-Linked Immunosorbent Assays (ELISAs). The traditional ELISA technique typically involves chromogenic reporters and substrates that produce some kind of visible color change to indicate the presence of an antigen or analyte. However, newer ELISA-like techniques use fluorogenic, electrochemiluminescent and quantitative polymerase chain reaction (PCR) reporters to create quantifiable signals. ELISA, as an optical approach for measurement, generally depended on bulky and power- intensive light sources, detectors, and monochromators; and suffered from potential false signals arising from complex colored samples. Moreover, because the sensitivity of optical methods follows the well-known Lambert-Beer law, a minimum sample volume and path length is required to achieve desired level of performance. In this context, electrochemical methods appear as a most promising alternative to optical approaches. The possibility to miniaturize modern microelectronics allows to build microelectrodes that are well suited for detection of very small volumes of samples (microliters to nanoliters). In fact, the sensitivity of electrochemical methods is not affected by the volume of samples used during the measurement. The low cost and a large-scale production of electronic devices is another reason that makes the electrochemical approach more appealing.

Test equipment based on electrochemical ELISA can be cost-effective point-of-care (POC) clinical diagnostic tools that can be deployed rapidly when needed and are useful in both, the developed world and in low resource settings such as semi-urban and rural areas in the developing countries. Most diseases or disorders including infectious, nutrient deficiency or lifestyle diseases could be easily detected using these tools thereby benefitting the human kind. There are innumerable diseases that plague humans, which if diagnosed early and accurately, could help to effectively treat the disease and can also avoid spreading of diseases particularly in case of infectious diseases.

Therefore, there is a need for a hand-held, point-of-care device for measurement of clinical diagnostic parameters using electrochemical ELISA (biosensor/immunosensor) platform technology.

SUMMARY OF THE INVENTION

Aspects of present invention relate to a system and a method for quantitative measurement of disease markers in biological samples. One should appreciate that although the present invention has been explained with reference to test for glycated hemoglobin (HbAlc), any other test for detection of disease markers such as cardiac markers, thyroid markers in a biological sample can be implemented through the proposed technique, all of which are entirely covered within the scope of the instant disclosure of the invention.

In an aspect, the present invention provides a sample analyzer device comprising an opening for receiving a cartridge, wherein the cartridge comprises one or more chambers for storing at least one reagent, a sample receiving port configured to receive a sample to be analyzed, and a test area having a sensor and a microfluidic channel. The sample analyzer device further includes at least one pump operatively coupled with the cartridge and configured to pump air into at least one of said one or more chambers for moving said at least one reagent to said test area. In another aspect, the present invention provides for a cartridge configured to enable testing of a sample; said cartridge comprising one or more chambers for storing at least one reagent, a sample receiving port configured to receive a sample to be analyzed, and a test area having a sensor and a microfluidic channel, wherein the cartridge is operatively coupled with a sample analyzer device, and wherein upon coupling of the cartridge with the sample analyzer device and initiation of the testing, means are enabled to allow flow of air into at least one of said one or more chambers to move the at least one reagent to the test area to enable reaction between the sample and the at least one reagent at the microfluidic channel.

In another aspect, the present invention relates to a method for analyzing a sample comprising the steps of: receiving a cartridge in an sample analyzer device, wherein the cartridge comprises one or more chambers for storing at least one reagent, a sample receiving port for receiving a sample to be analyzed, and a test area comprising a sensor and a microfluidic channel; receiving a sample to be analyzed on the sample receiving port of the cartridge; initiating test procedure for the sample, wherein the step of initiating starts at least one pump of the sample analyzer device, wherein said at least one pump pumps air into at least one of the one or more chambers for moving the at least one reagent to the test area for reaction between the at least one reagent and the sample in the microfluidic channel; and receiving signal from the sensor in the sample analyzer device to enable analysis of the signal to present test results.

In another, the present invention provides systems and methods for use of ELISA platform technology following highly reliable methodology of onboard reagent washing and reading, common to typical large laboratory instruments for rapid quantitative measurement of low concentration disease markers such as diabetic, cardiac, thyroid etc. in biological sample. In another aspect, the present invention provides systems and methods for use of electrochemical detection technique to provide a hand-held, point-of-care device, which is inexpensive, single use and having disposable microfluidic cartridge for rapid quantitative measurement of low concentration disease markers such as diabetic, cardiac, thyroid etc. in a biological sample.

In yet another aspect, the present invention provides various interface means such as printer, graphical touch screen display and USB (Universal Serial Bus) port and/or chip port to make wide variety of interface options available to the user to operate the device even in low resource settings. Various objects, features, aspects and advantages of the present invention will become more apparent from the following detailed description of preferred embodiments, along with the accompanying drawing figures, in which like numerals represent like components.

BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings are included to provide a further understanding of the present invention, and are incorporated in and constitute a part of this specification. The drawings illustrate exemplary embodiments of the present invention and, together with the description, serve to explain the principles of the present invention.

FIG. 1 illustrates exemplary functional modules of the system and methods for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention.

FIG. 2(a) illustrates a front perspective view of an exemplary analyzer for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. FIG. 2(b) illustrates a rear perspective view of an exemplary analyzer for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention.

FIG. 2(c) illustrates a perspective view indicating exemplary cartridge inserted through locator of exemplary analyzer for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. FIG. 3(a) illustrates a perspective view of interior of front part of exemplary analyzer for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention.

FIG. 3(b) illustrates a perspective view of interior of rear part of exemplary analyzer for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention.

FIG. 4(a) illustrates a perspective view of exemplary cartridge for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention.

FIG. 4(b) illustrates a perspective view indicating interior configuration of exemplary cartridge for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention.

FIG. 4(c) illustrates another perspective view indicating interior configuration including microfluidic channels of exemplary cartridge for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. FIG. 4(d) illustrates yet another perspective view indicating configuration of one way valve in the exemplary cartridge for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention.

FIG. 5(a) illustrates a schematic diagram of exemplary needle base assembly in accordance with embodiments of the present invention. FIG. 5(b) illustrates a schematic diagram indicating an exemplary assembly of needle base assembly with printed circuit board and connector in accordance with embodiments of the present invention.

FIG. 5(c) illustrates a schematic diagram indicating cartridge duly received in needle base assembly and connector in accordance with embodiments of the present invention. FIG. 6 illustrates a perspective view of interior of exemplary analyzer indicating configuration of pumps, needle base assembly and pipe connections between them in accordance with embodiments of the present invention.

FIG. 7 illustrates an exemplary process flow diagram for method of quantitative measurement of low concentration disease markers in biological samples in accordance with embodiments of present invention. FIG. 8 illustrates an exemplary process flow diagram for process of quantitative measurement of low concentration disease markers in biological samples in accordance with embodiments of the present invention.

FIG. 9(a) illustrates an exemplary screen shot of configuration page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(b) illustrates an exemplary screen shot of home page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(c) illustrates an exemplary screen shot of select test page of display screen of analyzer in accordance with embodiments of the present invention. FIG. 9(d) illustrates an exemplary screen shot of step-1 basic information page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(e) illustrates an exemplary screen shot of step-2 run test page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(f) illustrates an exemplary screen shot of insert cartridge page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(g) illustrates an exemplary screen shot of insert chip page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(h) illustrates an exemplary screen shot of blood sample page of display screen of analyzer in accordance with embodiments of the present invention. FIG. 9(i) illustrates an exemplary screen shot of processing page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(j) illustrates an exemplary screen shot of apply sample page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(k) illustrates an exemplary screen shot of close cap of cartridge page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(1) illustrates an exemplary screen shot of processing page of display screen of analyzer in accordance with embodiments of the present invention.

FIG. 9(m) illustrates an exemplary screen shot of result display page of display screen of analyzer in accordance with embodiments of the present invention. FIG. 9(n) illustrates an exemplary screen shot of data screen page of display screen of analyzer in accordance with embodiments of the present invention.

DETAILED DESCRIPTION OF THE INVENTION Before the present invention is described in further detail, it is to be understood that the detailed description and specific examples, while indicating embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art. One skilled in the art, based upon the description herein, can utilize the present invention to its fullest extent. Unless defined otherwise, all technical and specific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs.

Unless the context requires otherwise, throughout the specification and claims which follow, the word "comprise" and variations thereof, such as, "comprises" and "comprising" are to be construed in an open, inclusive sense that is as "including, but not limited to". The phrases "one embodiment" or "an embodiment" referred throughout the specification means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

As used in the specification and claims, singular terms including, but not limited to, "a", "an" and "the" include plural references unless the context clearly indicates otherwise. Plural terms include singular references unless the context clearly indicates otherwise.

It should also be noted that the term "or" is generally employed in its sense including "and/or" unless the content clearly dictates otherwise.

The term "biological sample" or "sample" as used herein refers to a sample of a body liquid obtained from a subject (e.g., a human). The said body liquid is selected from the group consisting of a blood sample, a urine sample and a saliva sample. The blood sample referred to herein encompasses a whole blood sample, a plasma sample or a serum sample. The term "biological sample" or "sample" can also include any material derived by processing the sample. Derived materials can include, but are not limited to, cells (or their progeny) isolated from the biological sample and proteins extracted from the sample. Processing of the biological sample can involve one or more of, filtration, distillation, extraction, concentration, fixation, inactivation of interfering components, addition of reagents, and the like.

The term "disease marker" as used herein refers to a substance; of which the concentration is altered, preferably elevated, in a biological sample from a diseased patient (subject) when compared to a normal healthy subject, and which can subsequently be used as a marker substance indicative of a disease. In the context of the present invention, the examples of disease markers include the markers for diabetes, cardiac, thyroid, and infectious diseases. The cardiac marker or the disease marker such as cardiac refers to markers/biomarkers such as cardiac troponin, creatine kinase-MB (CK-Mb), B-type natriuretic peptide (BNP) etc. The term "infectious diseases" as used herein refers to those diseases that are caused by pathogens including, but not limited to, viruses, bacteria, archaea, planaria, amoeba, and fungi.

Reference will now be made in detail to the exemplary embodiments of the present invention, examples of which are illustrated in the accompanying drawings. Wherever possible, the same reference numbers are used in the drawings and the description to refer to the same or like parts.

The following discussion provides several exemplary embodiments of the subject matter. Although each embodiment represents a single combination of the elements of the instant invention, the subject matter is considered to include all possible combinations of the disclosed elements. Thus, if one embodiment comprises elements A, B, and C, and a second embodiment comprises elements B and D, then the subject matter is also considered to include other remaining combinations of A, B, C, or D, even if not explicitly disclosed.

The embodiments of present invention describe systems and methods for quantitative measurement of disease markers such as diabetic, cardiac or thyroid in biological samples including, but not limited to, blood or serum, wherein device disclosed for testing can be hand- held and/or point-of-care device.

In an embodiment, the present invention provides systems and methods for use of electrochemical ELISA platform technology and can follow highly reliable methodology of onboard reagent washing and reading that can be common to typical large laboratory instruments at the same time miniaturizing laboratory set up to provide a hand-held, point-of-care device for testing various disease markers in biological samples.

In another embodiment, the present disclosure provides system and method for use of inexpensive, single use, disposable microfluidic cartridge for rapid quantitative measurement of analytes in a sample, wherein quantity of sample is very small, for instance in microliters. For example in case of blood sample, the blood can be obtained by pricking finger of patient thereby doing away with requirement of specialist to draw blood from human body using intravenous means and making process of sample collection simple and rapid. According to an aspect, the present invention provides a sample analyzer device comprising an opening for receiving a cartridge, wherein the cartridge comprises one or more chambers for storing at least one reagent, a sample receiving port configured to receive a sample to be analyzed, and a test area having a sensor and a microfluidic channel. The sample analyzer device can further include at least one pump operatively coupled with the cartridge and configured to pump air into at least one of said one or more chambers for moving said at least one reagent to said test area.

In another aspect, the present invention provides for a cartridge configured to enable testing of a sample; said cartridge comprising one or more chambers for storing at least one reagent, a sample receiving port configured to receive a sample to be analyzed, and a test area having a sensor and a microfluidic channel, wherein the cartridge is operatively coupled with a sample analyzer device, and wherein upon coupling of the cartridge with the sample analyzer device and initiation of the testing, means are enabled to allow flow of air into at least one of said one or more chambers to move the at least one reagent to the test area to enable reaction between the sample and the at least one reagent at the microfluidic channel. In yet another embodiment, the present invention provides a sample analyzer device having a means for pumping air to move reagents from reservoirs to test area to perform functions of a traditional ELISA analyzer, wherein such means can include one or more miniature pump(s) that can be configured to pump air to reagent reservoirs of a cartridge that stores reagents by pushing the reagent(s) to the test area in a predetermined sequence for replicating traditional ELISA process for testing of disease markers in biological samples.

In yet another embodiment, the present invention provides systems and methods enabling an interactive procedure for carrying out tests on a biological sample for various disease markers, wherein a user can be guided through various steps involved in the test method, thus doing away with any specialized training for user and simplifying and speeding up the process of testing of biological samples for various disease markers.

In a disclosure of the present invention, for ease of reference, the one or more chambers comprised in the cartridge can be referred to as first chamber, second chamber, third chamber, fourth chamber etc. Further, the chambers comprised in the cartridge can be operatively coupled with one another by means of a one way valve. For instance, the first chamber is operatively coupled with the second chamber by means of a one way valve.

FIG. 1 illustrates exemplary functional modules of system 100 for quantitative measurement of disease markers in biological samples such as blood/serum samples in accordance with embodiments of present disclosure. In an embodiment, system 100 can include a test cartridge module 102, and an analyzer module 104, wherein a means can first be incorporated to collect biological sample such as blood in small quantities, such as in microliters and can be configured to use a lancet for pricking finger of a patient where a generous drop of blood can be developed and can be collected by means of a pipet. In an alternate embodiment, there can be venous whole blood sample collected in a tube, which can be mixed by inverting the tube 8-10 times before collection in a pipet. One should appreciate that any other means for collecting the sample is well within the scope of the present disclosure and the implementations described herein are only exemplary in nature. In another embodiment, test cartridge module 104 can be configured as in a microfluidic device such as a cartridge having microfluidic channels configured to facilitate test process for biomarkers in small quantity of biological samples. The cartridge can also incorporate one or more chambers to store one or more reagents (hereinafter also referred to as reservoirs or like terms used interchangeably) required during the process of quantitative testing of disease markers in biological samples. The reservoirs can be in fluidic communication with microfluidic channels through one way valves that can permit movement of stored reagents from chambers to micro channels, but prevent their return back to chambers. The cartridge can additionally have sensors to pick up electric signals from electrochemical process, which can take place between the biological sample and one or more reagents. In an embodiment, the cartridge can be configured to receive a biological sample, which can travel to a test area within the micro channels and in fluidic communication with reagent reservoirs. In another embodiment, the cartridge can be configured to receive air in the reservoirs, which can propel reagents to test area. Thus, the test cartridge module 104 can be configured to replicate ELISA test platform similar to typical large laboratory instruments with electrochemical detection technique. In an embodiment there can be a different cartridge for testing of each of the specific disease markers, wherein reagents stored in reservoirs are specific to test requirements. In another embodiment test cartridge module 104 can also include a chip that is configured to carry calibration data pertaining to the cartridge. In an implementation, there can be one common chip for a batch of cartridges having similar calibration characteristics, wherein a cartridge can be used in combination with corresponding chip to make the test cartridge module 104 self calibrating.

In another embodiment of the present invention, an analyzer module 106 can be configured in an analysis point-of-care device, and can include a cartridge receiving module 108, a control module 110, a fluid movement module 112, a user interface module 114, and a power module 116, wherein the cartridge receiving module 108 can be configured to receive a test cartridge in a front side slot/opening and a chip in the back side slot/opening (details of these means are elaborated in succeeding paragraphs). In an implementation, the chip slot can have a means to put chip in electrical communication with analyzer module 106 so that calibration data stored in the chip can be available to analyzer module 106. Likewise, the slot/opening for receiving the cartridge can have a means to put sensors in microfluidic channels in electrical communication with analyzer module 106 so that signal from electrochemical process can be available to the analyzer module 106. In addition, cartridge receiving module 108 can be configured with a means to put cartridge reservoirs in fluidic communication with fluid movement module 112.

In an embodiment, control module 110 can be configured to control test process, interpret test data, and interface with the user of the analyzer device. In an embodiment, the module 110 can be configured to store various test procedures for different disease markers, give options to user to select one among the different tests, and implement relevant test procedure(s) based on selection(s) made by the user. The module 110 can also be configured to control fluid movement module 112 to facilitate movement of one or more reagents stored in reservoirs of test cartridge module 104 in predetermined sequence in accordance with the test procedure corresponding to the selected test. According to one embodiment, analyzer device of the present invention can include a means to pump air to reservoirs in the test cartridge to push reagents stored therein to microfluidic channels of the test cartridge. Such means can include a plurality of miniature pumps, for instance two pumps, including, but not limited to, peristaltic pumps. In an embodiment, there can be as many pumps as the number of reagent reservoirs each for pumping air to one reservoir. Working of these pumps can be controlled by the control module 110 in accordance with the desired test requirement. Fluid movement module 112 can also include means to bring these pumps in fluid connectivity with reservoirs as and when the test cartridge module 104 is received in the analyzer module 106, wherein details of these means are elaborated in succeeding paragraphs. In an embodiment, the control module 110 can also facilitate an interactive user interface through user interface module 114 to guide user through various steps of test procedure in a sequential manner, thus facilitating testing of disease markers by user without any detailed training. The user interface module 114 can include various modules including but not limited to graphical touch screen display, printer, USB port/chip port and other such modules. Such interface devices can be chosen so as to be suitable for hand-held, point-of-care devices, and at the same time not compromising quality of user interface. In an implementation, these modules can be controlled by the control module 110. Power module 116 can include a storage device such as rechargeable battery to power the analyzer module 106, a charging circuit, and power switch.

FIG. 2(a) illustrates perspective front view 200 of an exemplary analyzer of device for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. As shown, the sample analyzer device can include a front cover 202, a display 204 such as having a 3.5 inches graphical touch screen display, an oval cover 206 (or any other shape of opening) that can incorporate an opening for receiving a test cartridge, and a printer cover 208 for the printer such as a one inch thermal printer.

FIG. 2(b) illustrates an exemplary perspective rear view 250 of an exemplary analyzer of device for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. As shown, the analyzer device can include a bottom part 252, a back part 254, a power switch 256, USB ports/chip ports 258, and power port 260.

FIG. 2(c) illustrates an exemplary perspective view 280 of the cover 206 along with cartridge 400, and arrangement of needle base assembly 500 in the interior configured to receive and support distal end of cartridge 400.

FIG. 3(a) illustrates perspective interior view 300 of the front part of exemplary sample analyzer device for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. The device, as illustrated, can include a needle base assembly 500 placed behind the cover 206 (which can be of any desired/configured shape) and configured to receive distal end of the cartridge 400 (not shown in the Fig.). In an embodiment of present invention, the needle base assembly 500 can be configured to bring reservoirs of cartridge 400 in fluidic communication with pumps of the fluid movement module 112 (details given in subsequent paragraphs). The FIG. 3(a) also illustrates display printed circuit board (PCB) 302, which is placed behind display 204 and configured to control display functions as part of control module 110. FIG. 3(b) illustrates perspective interior view 350 of rear part of exemplary sample analyzer device for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. The device, as illustrated, can include a main PCB 352 mounted on bottom part 252, and a printer PCB 354 that can form part of the control module 110. The device can further include a printer 356 mounted on printer holder 362, a paper roll 358 of printer 356, and a paper roll holding part 360. There is also housed a pump A 364 and a pump B 366 and both these pumps form part of fluidic movement module 112. The pumps are miniature pumps such as, but not limited to, peristaltic pumps, which are configured to pump air to reservoirs of cartridge 400 to move reagents to test area. In an embodiment, operation of these pumps can be controlled by control module 110 in a predetermined manner in accordance with test requirement.

FIG. 4(a) illustrates perspective view of an exemplary cartridge 400 for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. The cartridge 400 can include a multichannel body 402 having a side cover 430 on each side, a port 428 for receiving biological sample, and a cover 426 for port 428 to close the port 428 after biological sample is received. It can also have microfluidic laminates 418 that can be configured to define microfluidic channels for flow of sample and reagents and test area. Test area can include a sensor (not shown) for detecting signal from electrochemical reaction. FIG. 4(b) illustrates another view of exemplary cartridge 400 for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. The view displays internal construction from right side of the cartridge 400 illustrating the first chamber 404 and the second chamber 406. Also illustrated are septum 422, which can cover and seal openings to first chamber 404 and third chamber 410 (described in succeeding paragraph). The septum 422 can be made of a suitable material including, but not limited to, silicon rubber, which can be punctured and penetrated by sharp and thin objects such as needles at the same time sealing the space around needles to prevent leakage of fluid stored in the chambers. These septum 422 can be configured to facilitate fluid connectivity of first chamber 404 and third chamber 410 with pump A 364 and pump B 366 respectively (detailed description in succeeding paragraphs)

FIG. 4(c) illustrates yet another view of exemplary cartridge 400 for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. The view illustrates internal construction from left side of the cartridge 400 illustrating the third chamber 410, the fourth chamber 408, and a waste chamber 412. Also illustrated are details of microfluidic laminate 418, wherein gate 420 can connect microfluidic channel with sample port 428, gate 414 can connect microfluidic channel to fourth chamber 408, gate 416 can connect microfluidic channel to second chamber 406, and gate 426 can connect microfluidic channel to waste chamber 412. The first chamber containing first reagent is moved from the first chamber to the second chamber before moving onto test area. The first reagent may comprises antibodies for binding to analyte of biological sample

FIG. 4(d) illustrates another view of exemplary cartridge 400 for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. The view illustrates configuration of one way valves 424 between third chamber 410 and fourth chamber 408. One way valve 424 can permit one way fluidic movement from third chamber 410 to fourth chamber 408 and can block fluidic connectivity if fluid were to move from fourth chamber 408 to third chamber 410. The third chamber containing second reagent is moved from the third chamber to the fourth chamber before moving onto test area. The second reagent may comprise at least one marker for the measurement of analyte of biological sample. There can be two one way valves 424, one between fourth chamber 408 and third chamber 410, and another one between second chamber 406 and first chamber 404.

FIG. 5(a) illustrates an exemplary needle base assembly 500 for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. The assembly 500 can include a needle base 502 that can incorporate two supporting pads 512 configured to support cartridge 400 (not shown in the Fig.) when received. The assembly 500 can further incorporate two hollow needles, namely needle A 504 and needle B 506 configured to puncture and penetrate septum(s) 422 of cartridge 400. In an embodiment, the hollow needles 504 and 506 on penetrating septum 422 can enter the first chamber 404 and third chamber 410 respectively, of cartridge 400, and can bring these chambers in fluidic communication with pump A 364 and pump B 366 by means (for instance PCB and/or connector), which is described in subsequent paragraphs.

FIG. 5(b) illustrates an exemplary configuration 550 of needle base assembly 500 with PCB 554 and connector 552. The connector 552 can bring cartridge 400 in electric communication with PCB 554 when cartridge is received in needle base assembly 500 and thus can enable transfer of electrical signal generated by sensor of cartridge 400. FIG. 5(c) illustrates an exemplary embodiment 580 of cartridge 400 duly received in needle base assembly 500 and supported by support pads 512. In this configuration, needles 504 and 506 can penetrate septum 422 and enter cambers 404 and 410 respectively.

FIG. 6 illustrates another interior view 600 of the exemplary sample analyzer device for quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. As shown, the sample analyzer device provides means to achieve fluidic communication between pumps 364 and 366 with needles 504 and 506 (not shown here) of needle base assembly 500. Tube A 608 and tube B 610 can connect output sides of pump A 364 and pump B 366 with needle A 504 and needle B 506 respectively to enable transfer of pumped air to first chamber 404 and third chamber 410 through needle A 504 and needle B 506. Also the figure illustrates a locator 606, which can facilitate mounting of needle base assembly 500 and PCB 554 on cover 206 (not shown here). There can be a plurality of locators 606 depending on configuration of mounting and assembly requirements. Locator(s) 606 can additionally guide cartridge 400 from point of its entry in opening in cover 206 and its safe positioning in needle base assembly 500 and can be configured to ensure that cartridge 400's location with reference to needles 504 and 506 and connector 552 is correct.

In an embodiment of the present invention, cartridge 400 can be inserted in the opening of the cover 206, wherein, guided by the locater 606, distal end of the cartridge 400 can reach needle base assembly 500 and can get supported by support pads 512. Simultaneously connector 552, which can protrude out of PCB 554 can come in contact with cartridge 400, thus providing electrical connectivity between cartridge 400 and PCB 554. During the insertion process of cartridge 400, needles 504 and 506 can puncture and penetrate septum 422 and enter chambers 404 and 410 respectively, creating positive connection between pumps 364 and 366 and chambers 404 and 410 respectively. After insertion of the cartridge 400 and receiving of biological sample in port 428, test cycle can be started. After start of the test cycle and requisite incubation period during which biological sample travels from port 428 to microfluidic laminate area 418 through gate 420, control module 110 can start pump A 364 and air can travel through tube A 608 and needle A 504 to first chamber 404 and can create pressure on stored reagent to open corresponding one way valve 424 and travel to second chamber 406, and thereafter to microfluidic laminate area 418 through gate 416. During this transfer process, pressure of the reagent removes trapped air from second chamber 406 and microfluidic laminate area 418 and move it to waste chamber 412. After predetermined time period, pump A 364 can stop, and after due incubation period, pump B 366 can start and through the corresponding tube and needle, pump air to third chamber 410, thereby transferring stored reagent to fourth chamber 408. Again, pressure of the reagent can move trapped air and liquid waste to waste chamber 412 and reagent can reach microfluidic laminate area 418. After specific/desired period of time, the pump B 366 stops. When the biological sample and reagents meet at microfluidic laminate area 418, desired reaction can take place between the sample and the reagents and the result can be calculated based on preset equation and can be displayed/printed at user interface.

FIG. 7 illustrates an exemplary process flow diagram 700 for a method of quantitative measurement of disease markers in biological samples in accordance with embodiments of the present invention. At step 702, the biological sample such as blood can be collected. For example, a lancet can be used to prick finger of patient and develop a drop of blood. There can be a capillary device such as a blood key with a dispensing tool at the end to collect the blood from the drop. Blood can be drawn into the key by touching end of the drop. There can be a lysing buffer vial containing suitable reagent for lysing the blood sample. The collected sample can be lysed by inserting sample key into the vial and shaking for sufficient time say 30 seconds after locking cap of the vial. At next step 704, the test cartridge can be received by sample analyzer device. In an embodiment, cartridge can be a microfluidic device configured to test very small quantities of biological samples and can be configured to carry reagents required for testing the disease markers. It can also include a sensor to pick up signal from electrochemical reaction between the biological sample and the reagents. Thus, the cartridge can replicate ELISA platform technology and can follow highly reliable methodology of onboard reagent washing & reading that can be common to typical large laboratory instruments. There can be different cartridges for different disease markers, wherein reagents stored in the cartridge are specific to the disease marker. One of the reagents can be conjugate buffer having a conjugated antibody and stabilizer, preservative and buffer. Other reagent can be a substrate buffer comprising substrate, surfactant, preservative and buffer. In addition, there can be a chip carrying calibration data pertaining to the cartridge and can also be received by the analyzer.

After the cartridge is received in the analyzer, at next step 706, lysed sample, collected at step 702, can be received at cartridge 400. In an embodiment, the biological sample is received at a port 428 disposed in the cartridge and the sample can travel to microfluidic laminate area. After the biological sample is received at cartridge, test cycle can be initiated at next step

708. In an embodiment, control module of analyzer can be configured to carry out test cycle without human intervention for carrying out the process in predetermined manner. At step 710, the control module can actuate pumps in predetermined sequence to move reagents stored in the cartridge to microfluidic laminate area in accordance with requirement of the test cycle. For example, the control module can first move conjugate buffer containing conjugated antibody to microfluidic area so as to allow attachment of antibodies to disease marker present in the biological sample and subsequently, after requisite incubation period, the substrate buffer moved to the microfluidic substrate area so that desired reaction can happen.

At step 712, on completion of reaction, electrochemical signal can be picked up by sensor configured on the cartridge and transferred to control module.

At step 714, signal received from the sensor can be analyzed and interpreted based on preset equation and calibration data pertaining to the cartridge.

At step 716, calculated result(s) can be displayed through user interface, which can be, for example, but not limited to, display screen or a printer.

In another aspect, the present invention relates to a method for analyzing a sample comprising the steps of receiving a cartridge in an sample analyzer device, wherein the cartridge comprises one or more chambers for storing at least one reagent, a sample receiving port for receiving a sample to be analyzed, and a test area comprising a sensor and a microfluidic channel; receiving a sample to be analyzed on the sample receiving port of the cartridge; initiating test procedure for the sample, wherein the step of initiating starts at least one pump of the sample analyzer device, wherein said at least one pump pumps air into at least one of the one or more chambers for moving the at least one reagent to the test area for reaction between the at least one reagent and the sample in the microfluidic channel; and receiving signal from the sensor in the sample analyzer device to enable analysis of the signal to present test results.

FIG. 8 illustrates an exemplary flow diagram 800 illustrating procedure of quantitative measurement of disease markers in biological samples in accordance with embodiments of present invention. At step 802 a user can put the power switch on, to energize the device. In accordance with an embodiment, powering the sample analyzer device can provide the user with an interactive interface for carrying out the test for disease marker in biological sample, wherein the user can be presented with a display on display screen an exemplary screen shot of which is provided at representation 900 of FIG. 9(a). The exemplary screen shot shows various configuration data pertaining to the device for example position pertaining to cartridge slot, chip slot, battery, printer paper and ambient temperature etc. It can also show details pertaining to reagents that are pumped by pumps A and B and are configured to move to microfluidic laminate area. The screen can also incorporate a button for moving to the next page, which can be home page, an exemplary illustration of which is provided at representation 905 of FIG. 9(b). Home page can, besides providing various options for setting the device, incorporate a test button that opens the next screen, an exemplary illustration of which is provided at representation 910 of FIG. 9(c).

At step 804, the user can select test to be performed using various options provided at exemplary display screen shot illustrated at FIG. 9(c). The display can provide options for testing different disease markers such as but not limited to cardiac, thyroid, diabetes etc. The user can click appropriate button upon which next screen, an exemplary illustration shown at representation 915 of FIG. 9(d), can be displayed which can ask for basic information such as operator ID and patient ID.

At step 806, the user can enter details pertaining to patient and user/operator and move to next screen which can be 'run test' screen, an exemplary illustration provided at representation 920 of FIG. 9(e). The screen can list various steps involved in running the test and accordingly apprise a new user about the sequence of steps, so that the new user is ready accordingly. Next screen can be "Step 1: Insert Cartridge" screen, an exemplary illustration provided at representation 925 of FIG. 9(f).

At step 808, in accordance with Step 1 displayed on display screen, the user can insert cartridge corresponding to test being carried out in slot provided in cover of device and can move to next screen which can be "Step 2: Insert Chip Screen", an exemplary illustration provided at representation 930 of FIG. 9(g).

At step 810, in accordance with step 2 displayed on display screen, the user can insert chip representing cartridge in chip slot and can move to next screen which can be "Step 3: Blood Sample", an exemplary illustration provided at representation 935 of FIG. 9(h). At step 812, the user can collect biological sample, for example, blood sample from a patient and thereafter pre-process it. In accordance with step 3 displayed on display screen, the user can prick patient's finger using lancet after wiping finger with alcohol and allowing it to dry. The user such as a pathologist or a doctor can develop a generous drop of blood, which can be collected using a pipet like capillary device such as blood key with dispensing tool which when kept at edge of the blood drop can suck the blood. User can also take precautions to ensure that the blood key is completely filled; no blood gets on the outside of the blood key and no air bubbles get into the key. The user can also ensure not to wipe off the blood key. User can thereafter move to the next screen, which can be "Step 4: Pre-processing Window", an exemplary illustration provided at representation 940 of FIG. 9(i). At this stage, the user can insert the blood key into lysing buffer vial and shake well for appropriate time for example 30 seconds, whereby sample can be ready for further processing. In an alternate scenario, venous whole blood sample can be collected in tubes in which case, sample can be mixed well by inverting the tube 8-10 times before pre-processing. User can now move to next screen, which can be "Step 5: Apply Sample", an exemplary illustration provided at representation 945 of FIG. 9(j).

At step 814, in accordance with step 5 displayed on display screen, the user can dispense biological sample such as blood collected and preprocessed at sample port of cartridge. The user can use dispensing tool of the blood key to discharge sample at the sample port. The user may be required to apply specified quantity of sample say for example 150 μΐ, which the user can control using markings provided on blood key. After applying sample, the user can move to next screen, which can be "Step 6: Close Cap of Cartridge", an exemplary illustration provided at representation 950 of FIG. 9 (k). User can close sample port with cap and move to next screen which can be "Step 2: Run Test", which is same as earlier illustrated at FIG. 9(e).

At step 816, the user can click on process button to start the test process. User can take precaution not to delay the start of process as it may lead to incorrect results. In an alternate embodiment, the user can directly start test process after insertion of cartridge, chip and application of biological sample without going through various screens described above, which are to guide the user through various steps of test procedure. After start of the test process, it can take some time for process to complete. The ELISA based test can take 7 to 15 minutes time to complete the process and show results. The enzyme assay based test can take less than 1 minute to complete the process and show the results. For example, a test for HbAlc can take 7 minutes and 30 seconds. Progress of test can be displayed on screen through a bar indicating percent of process completed. Exemplary screen shots are provided at representation 955 of FIG. 9(1) and representation 960 of FIG. 9(m). The screen can also display stage of process such as sample incubation, conjugate incubation and analyzing. If required the test process can be terminated at any stage by clicking the cancel button. After the test is completed the result can be displayed on screen. An exemplary screen shot with test results is illustrated at representation 965 of FIG. 9(n).

At step 818, the user can either record result from display screen or use option to print the result by using print option provided on screen as print button. Any other option can be configured to enable the user to take a desired action and therefore all such actions are completely within the scope of the present invention. On clicking the print button, printer provided on the device can print test result along with other relevant details such as patient's ID, operator's ID etc. At the next and last step 820, the user can dispose the used cartridge, blood key and lysing reagent vial in biological hazard waste container.

Thus, embodiments of present disclosure provide systems and method for low cost, sensitive and rapid measurement of disease markers such as diabetic, cardiac, thyroid markers and infectious diseases in low resource settings such as in semi-urban and rural areas in the developing countries using lab-on-cartridge based electrochemical ELISA platform technology that will expedite patient's treatment and provide overall improvement in the outcomes, including enhanced physician and patient satisfaction.

The above description represents merely an exemplary embodiment of the present invention, without any intention to limit the scope of the present invention thereto. Various equivalent changes, alterations or modification based on the present invention are all consequently viewed as being embraced by the scope of the present invention.

As used herein, and unless the context dictates otherwise, the term "coupled to" is intended to include both direct coupling (in which two elements that are coupled to each other contact each other) and indirect coupling (in which at least one additional element is located between the two elements). Therefore, the terms "coupled to" and "coupled with" are used synonymously. Within the context of this document terms "coupled to" and "coupled with" are also used euphemistically to mean "communicatively coupled with" over a network, where two or more devices are able to exchange data with each other over the network, possibly via one or more intermediary device. It should be apparent to those skilled in the art that many more modifications besides those already described are possible without departing from the inventive concepts herein. The inventive subject matter, therefore, is not to be restricted except in the spirit of the appended claims. Moreover, in interpreting both the specification and the claims, all terms should be interpreted in the broadest possible manner consistent with the context. In particular, the terms "comprises" and "comprising" should be interpreted as referring to elements, components, or steps in a non-exclusive manner, indicating that the referenced elements, components or steps may be present, or utilized, or combined with other elements, components, or steps that are not expressly referenced.

Claims

CLAIMS We claim -

1. A sample analyzer device comprising: a) an opening for receiving a cartridge, said cartridge comprising:

(i) one or more chambers for storing at least one reagent,

(ii) a sample receiving port configured to receive a sample to be analyzed, and

(iii) a test area comprising a sensor and a microfluidic channel; b) at least one pump operatively coupled with said cartridge and configured to pump air into at least one of said one or more chambers for moving said at least one reagent to said test area.

2. The device as claimed in claim 1, wherein said cartridge comprises a first chamber having a first reagent, wherein said first reagent is moved from said first chamber to a second chamber before moving onto said test area, wherein said first reagent comprises antibodies for binding to analyte of said sample.

3. The device as claimed in claim 2, wherein said first chamber is operatively coupled with said second chamber by means of a one way valve.

4. The device as claimed in claim 1, wherein said cartridge comprises a third chamber having a second reagent, wherein said second reagent is moved from said third chamber to a fourth chamber before moving onto said test area, wherein said second reagent comprises at least one marker for measurement of analyte of said sample.

5. The device as claimed in claim 4, wherein said third chamber is operatively coupled with said fourth chamber by means of a one way valve.

6. The device as claimed in claim 1, wherein said cartridge comprises a waste chamber configured to collect air and liquid waste.

7. The device as claimed in claim 1, wherein said at least one pump is operatively coupled with a means for communicating with said cartridge comprising one or more chambers so as to move air from said at least one pump to said one or more chambers.

8. The device as claimed in claim 7, wherein said means are needles.

9. The device as claimed in claim 7, wherein said means puncture and penetrate at least one septum, wherein said at least one septum is configured to cover said one or more chambers such that when said cartridge is inserted in said device, said needles are actuated to puncture and penetrate said at least one septum to make air from said at least one pump to flow through to said one or more chambers.

10. The device as claimed in claim 7, wherein said device comprises two miniature pumps.

11. The device as claimed in claim 10, wherein said miniature pumps are peristaltic pumps.

12. The device as claimed in claim 1, wherein said device comprises a display screen to enable a user of said device to operate said device.

13. The device as claimed in claim 12, wherein said display means is touch-screen enabled.

14. The device as claimed in claim 1, wherein said device comprises a printer.

15. The device as claimed in claim 1, wherein said device is operatively coupled with a battery to enable operation of said device.

16. The device as claimed in claim 1, wherein said sample comprises a blood sample or a serum sample.

17. The device as claimed in claim 1, wherein said device is configured to analyze one or more disease markers for diseases comprising diabetes, cardiac, thyroid, and infectious diseases.

18. A method for analyzing a sample comprising: a) receiving a cartridge in an analyzer device, wherein said cartridge comprises one or more chambers for storing at least one reagent, a sample receiving port for receiving a sample to be analyzed, and a test area comprising a sensor and a microfluidic channel; b) receiving a sample to be analyzed on said sample receiving port of said cartridge; c) initiating test procedure for said sample, wherein said step of initiating starts at least one pump of said analyzer device, wherein said at least one pump pumps air into at least one of said one or more chambers for moving said at least one reagent to said test area for reaction between said at least one reagent and said sample in said microfluidic channel; and d) receiving signal from said sensor in said analyzer device to enable analysis of said signal to present test results.

19. The method as claimed in claim 18, wherein said cartridge comprises a first chamber having a first reagent, wherein said first reagent is moved from said first chamber to a second chamber before moving onto said test area, wherein said first reagent comprises antibodies for binding to analyte of said sample.

20. The method as claimed in claim 19, wherein said first chamber is operatively coupled with said second chamber by means of a one way valve.

21. The method as claimed in claim 18, wherein said cartridge comprises a third chamber having a second reagent, wherein said second reagent is moved from said third chamber to a fourth chamber before moving onto said test area, wherein said second reagent comprises at least one marker for measurement of analyte of said sample.

22. The method as claimed in claim 21, wherein said third chamber is operatively coupled with said fourth chamber by means of a one way valve.

The method as claimed in claim 18, wherein said cartridge comprises a waste chamber configured to collect air and liquid waste.

24. The method as claimed in claim 18, wherein said at least one pump is operatively coupled with a means for communicating with said one or more chambers so as to move air from said at least one pump to said one or more chambers.

The method as claimed in claim 24, wherein said means are needles.

26. The method as claimed in claim 24, wherein said means puncture and penetrate at least one septum, wherein said at least one septum is configured to cover said one or more chambers such that when said cartridge is inserted in said device, said needles are actuated to puncture and penetrate said at least one septum to make air from said at least one pump to flow through to said one or more chambers.

27. The method as claimed in claim 24, wherein said device comprises two miniature pumps.

28. The method as claimed in claim 24, wherein said miniature pumps are peristaltic pumps.

29. The method as claimed in claim 18, wherein said sample comprises a blood sample or a serum sample.

30. The method as claimed in claim 18, wherein said device is configured to analyze one or more disease markers for diseases comprising diabetes, cardiac, thyroid, and infectious diseases.

31. A cartridge configured to enable testing of a sample comprising a) one or more chambers for storing at least one reagent; b) a sample receiving port configured to receive a sample to be analyzed; and c) a test area comprising a sensor and a microfluidic channel, wherein said cartridge is operatively coupled with an analyzer device, and wherein upon coupling of said cartridge with said analyzer device and initiation of said testing, means are enabled to allow flow of air into at least one of said one or more chambers to move said at least one reagent to said test area to enable reaction between said sample and said at least one reagent at said microfluidic channel.

32. The cartridge as claimed in claim 31, wherein said means comprise at least one pump and one or more members such that upon initiation of said testing, said at least one pump enables said flow of air into at least one of said one or more chambers through said one or more members.

33. The cartridge as claimed in claim 32, wherein said one or more members comprise needles.

34. The cartridge as claimed in claim 31, wherein said cartridge is electrically coupled with said sample analyzer device.