JP2005265629A - Card type target material detector - Google Patents

Card type target material detector Download PDF

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JP2005265629A
JP2005265629A JP2004079034A JP2004079034A JP2005265629A JP 2005265629 A JP2005265629 A JP 2005265629A JP 2004079034 A JP2004079034 A JP 2004079034A JP 2004079034 A JP2004079034 A JP 2004079034A JP 2005265629 A JP2005265629 A JP 2005265629A
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electrode
card
temperature
target substance
type target
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JP4136979B2 (en
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Manabu Harada
学 原田
Tadashi Matsunaga
是 松永
Takeshi Tanaka
剛 田中
Taikei Hayashi
泰圭 林
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Tokyo University of Agriculture and Technology NUC
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Abstract

<P>PROBLEM TO BE SOLVED: To provide a card type target material detector capable of conducting accurate measurement. <P>SOLUTION: In the card type target material detector having a recessed part 1 for loading one or more than two sample(s) and an electrode 2 for electrochemically detecting a target material on a card type board, the recessed part 1 and the electrode 2 are connected with a flow passage having capillary force. The card type target material detector is provided with a temperature detector for detecting temperature of sample liquid moving to the electrode and a temperature adjuster for controlling the temperature of the sample liquid moving to the electrode. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

本発明は、正確に測定可能な、電気化学的手段により標的物質を検出するカード型標的物質検出装置に関する。   The present invention relates to a card-type target substance detection apparatus that detects a target substance by electrochemical means that can be accurately measured.

近年、マイクロマシーニングにより化学分析システムを小型化する研究が盛んに行われている。これは、システムの小型化により、無効体積の減少、サンプルや試薬量の低減、システムの低価格化、使用時の低消費電力化が図れるためである。システムの小型化の手段として、そのシステムを構成する個々の要素を小型化することが行われており、例えば、心電位測定用の体表面装着型マイクロシステムといったものが知られている。また、ポイント・オブ・ケア（ＰＯＣ）検査のために電極反応物と電極との電極反応を利用した電気化学的検出技術を導入したマイクロシステム（特許文献１）が研究され、小型化・低価格化が実現可能な段階にまで来ている。これらの電気化学的手法を用いたマイクロシステムは反応物質の濃度変化を電流値として示すものである。
特開２００３−９８１７１号公報 In recent years, research on reducing the size of chemical analysis systems by micromachining has been actively conducted. This is because the size of the system can be reduced, thereby reducing the ineffective volume, reducing the amount of samples and reagents, reducing the cost of the system, and reducing power consumption during use. As a means for downsizing the system, individual elements constituting the system are downsized. For example, a body surface-mounted microsystem for measuring an electrocardiogram is known. In addition, a microsystem (Patent Document 1) that introduces an electrochemical detection technique using an electrode reaction between an electrode reactant and an electrode for point-of-care (POC) inspection has been studied, and has been reduced in size and price. Has reached the stage where it can be realized. Microsystems using these electrochemical techniques show changes in the concentration of reactants as current values.
JP 2003-98171 A

しかしながら、当該マイクロシステムにおいては、測定値の誤差が大きく、特に医療分野において病気の診断基準として用いられるので１％以下の測定精度が求められ、当該システムの改良が望まれている。
従って、本発明の目的は、より正確な測定が可能なカード型標的物質検出装置を提供することにある。 However, the micro system has a large error in measurement values, and is used as a diagnostic standard for diseases particularly in the medical field. Therefore, a measurement accuracy of 1% or less is required, and improvement of the system is desired.
Accordingly, an object of the present invention is to provide a card type target substance detection apparatus capable of more accurate measurement.

そこで本発明者は、当該マイクロシステムにおける誤差の発生原因について種々検討したところ、マイクロシステムにおいては密閉された微細流路中で反応を行い、標的物質の電流値を測定するものであるが、反応が密閉された微細流路中で行われるため試料液の温度が上昇してしまうことが多く、その温度上昇に伴ない電流値が変化してしまい、測定値に誤差が生じてしまうものであることを見出した。そして、カード型標的物質検出装置に、温度検出器と温度調節器とを配置するか、又は温度検出器とその検出された温度変化に基づいて測定値を補正するための測定値制御部とを配置すれば、正確な測定値が得られることを見出し、本発明を完成した。   Therefore, the present inventor has studied various causes of the error in the microsystem, and in the microsystem, the reaction is performed in a closed fine channel and the current value of the target substance is measured. Since the temperature of the sample solution is often increased in a closed fine channel, the current value changes with the temperature increase, resulting in an error in the measured value. I found out. Then, a temperature detector and a temperature controller are arranged in the card-type target substance detection device, or a temperature detector and a measurement value control unit for correcting the measurement value based on the detected temperature change are provided. As a result, it was found that accurate measurement values can be obtained if arranged, and the present invention was completed.

すなわち、本発明は、カード型基板上に、（１）１又は２以上の試料載置用凹部、及び（２）標的物質を電気化学的に検出するための電極を有し、当該（１）凹部及び（２）電極が毛細管力を有する流路で連結されてなるカード型標的物質検出装置において、前記電極に移動する試料液の温度を検出するための温度検出器、及び前記電極に移動する試料液の温度を制御するための温度調節器が配置されていることを特徴とするカード型標的物質検出装置を提供するものである。   That is, the present invention has (1) one or two or more sample mounting recesses and (2) an electrode for electrochemically detecting a target substance on a card-type substrate, In a card-type target substance detection device in which a recess and (2) an electrode are connected by a flow path having a capillary force, a temperature detector for detecting the temperature of a sample solution that moves to the electrode, and the electrode moves to the electrode The present invention provides a card type target substance detection device in which a temperature controller for controlling the temperature of a sample solution is arranged.

また、本発明は、カード型基板上に、（１）１又は２以上の試料載置用凹部、及び（２）標的物質を電気化学的に検出するための電極を有し、当該（１）凹部及び（２）電極が毛細管力を有する流路で連結されてなるカード型標的物質検出装置において、前記電極に移動する試料液の温度を検出するための温度検出器、及び前記温度検出器により検出された温度に基づいて電気化学的に検出された測定値を補正するための測定値制御部が配置されていることを特徴とするカード型標的物質検出装置を提供するものである。   Moreover, this invention has (1) one or two or more sample mounting recessed parts on a card-type board | substrate, and (2) the electrode for detecting a target substance electrochemically, The said (1) In the card-type target substance detection device in which the recess and the electrode are connected by a flow path having a capillary force, a temperature detector for detecting the temperature of the sample liquid moving to the electrode, and the temperature detector The present invention provides a card type target substance detection device in which a measurement value control unit for correcting a measurement value detected electrochemically based on a detected temperature is arranged.

本発明の装置を用いれば、微細流路中で試料液の温度が変化しても正確な測定値が得られる。   By using the apparatus of the present invention, an accurate measurement value can be obtained even if the temperature of the sample solution changes in the fine flow path.

本発明のカード型標的物質検出装置は、カード型基板上に、（１）１又は２以上の試料載置用凹部、及び（２）標的物質を電気化学的に検出するための電極を有し、該（１）凹部及び（２）電極が毛細管力を有する流路で連結されてなるものである。当該カード型基板上には、さらに１又は２以上の試料移動用液体載置用凹部が設けられていてもよい。図１には、本発明カード型標的物質検出装置の一例である、（１）試料載置用凹部が１個、（２）液体載置用凹部が２個の場合の概念図を示してある。   The card-type target substance detection apparatus of the present invention has (1) one or two or more sample placement recesses on a card-type substrate, and (2) an electrode for electrochemically detecting the target substance. The (1) recess and (2) electrode are connected by a flow path having a capillary force. On the card-type substrate, one or two or more sample placement liquid placement recesses may be provided. FIG. 1 shows an example of the card-type target substance detection device according to the present invention, where (1) one sample mounting recess and (2) two liquid mounting recesses are conceptual diagrams. .

カード型基板は、合成樹脂製であるのが好ましく、当該合成樹脂としてはポリエチレン、ポリプロピレン、ポリメチル（メタ）アクリレート等が挙げられるが、特にポリメチル（メタ）アクリレートが好ましい。カード型基板は、例えば縦３０〜８０mm、横６０〜１５０mm、厚さ１〜３mm程度の板状物であるのが好ましい。   The card-type substrate is preferably made of a synthetic resin, and examples of the synthetic resin include polyethylene, polypropylene, polymethyl (meth) acrylate, and the like, and polymethyl (meth) acrylate is particularly preferable. The card-type substrate is preferably a plate-like material having a length of 30 to 80 mm, a width of 60 to 150 mm, and a thickness of 1 to 3 mm, for example.

（１）試料載置用凹部は１個でもよく、２個以上あってもよい。当該凹部は、試料が、反応に必要な量載置できる大きさであればよい。なお、試料移動用液体載置用凹部は、試料が液体、例えば試料含有溶液である場合には、特に必要とされない。試料や試料移動用液体は、通常、当該凹部にピペットにより滴下することにより載置される。なお、試料移動用液体としては、反応溶媒、緩衝液、水等が用いられる。 (1) The number of the sample mounting recesses may be one, or two or more. The recess may be of a size that allows the sample to be placed in an amount necessary for the reaction. Note that the sample moving liquid mounting recess is not particularly required when the sample is a liquid, for example, a sample-containing solution. The sample and the liquid for moving the sample are usually placed by dropping them into the concave portion with a pipette. As the sample moving liquid, a reaction solvent, a buffer solution, water, or the like is used.

（２）電極は、標的物質を電気化学的に検出できる構成であればよいが、電気化学的な方法による反応液の測定は、サイクリックボルタンメトリー（CV）がよく利用される。本法は反応液中にある３電極（参照電極、作用電極、対電極）において参照電極に対する作用電極の電位を変化させることで得られる酸化電流、還元電流の変化を読み取るものである。従って、当該電極は、例えば図２のように配置される。 (2) The electrode may be configured so that the target substance can be detected electrochemically, but cyclic voltammetry (CV) is often used for measuring the reaction solution by an electrochemical method. This method reads changes in oxidation current and reduction current obtained by changing the potential of the working electrode with respect to the reference electrode in three electrodes (reference electrode, working electrode, counter electrode) in the reaction solution. Accordingly, the electrodes are arranged as shown in FIG. 2, for example.

前記（１）試料載置用凹部、（２）電極及び必要に応じて試料移動用液体装置凹部は、毛細管力を有する流路で連結されている。流路は、例えば、カード型基板上に溝を掘ることによって形成することができ、流路の毛細管力は、溝の大きさ（幅）によって調整することができる。これは、溝の幅を小さくすれば毛細管力は大きくなり、溝の幅を大きくすれば毛細管力は小さくなる。流路のパターンは特に制限されない。   The (1) sample mounting recess, (2) the electrode, and optionally the sample moving liquid device recess are connected by a flow path having a capillary force. The flow path can be formed, for example, by digging a groove on a card-type substrate, and the capillary force of the flow path can be adjusted by the size (width) of the groove. This is because the capillary force increases as the groove width decreases, and the capillary force decreases as the groove width increases. The pattern of the flow path is not particularly limited.

このように試料載置用凹部と電極とが毛細管力を有する流路で連結されることにより、試料載置用凹部に滴下された試料液は流路を移動し、流路中で必要な反応を生起し、電極により標的物質の量が電気化学的に、電流として検出される。得られた電流値を用い、予め作成された検量線などから、標的物質の量を定量できる。   In this way, the sample mounting recess and the electrode are connected by a flow path having a capillary force, so that the sample liquid dropped on the sample mounting recess moves through the flow path, and the necessary reaction in the flow path. The amount of the target substance is detected electrochemically as an electric current by the electrode. Using the obtained current value, the amount of the target substance can be quantified from a calibration curve prepared in advance.

この電流は液中の標的物質の電極への拡散速度に比例する。また拡散速度は物質の濃度勾配に比例する。
従って電流ｉは、式（１） This current is proportional to the diffusion rate of the target substance in the liquid to the electrode. The diffusion rate is proportional to the concentration gradient of the substance.
Therefore, the current i is given by the formula (1)

Figure 2005265629
Figure 2005265629

ｎ：反応電子数 Ｆ：ファラデー定数（C/mol）
Ａ：電極面積（m2） Ｄ：反応液の拡散係数（m2/S）
Ｃ：反応液濃度（mol/m3） ｘ：電極表面から溶液内部への距離（m） n: Number of reaction electrons F: Faraday constant (C / mol)
A: Electrode area (m 2 ) D: Diffusion coefficient of reaction solution (m 2 / S)
C: Reaction solution concentration (mol / m 3 ) x: Distance from electrode surface to solution inside (m)

（１）式における拡散係数Ｄは水溶液系ならおよそ１０-9 m2/S位であるが、問題はDは温度依存性がある事である。拡散係数は液の粘度によって上下し、粘度は温度によって上下するからである。
従って標的物質の濃度を電流によって判定する場合、液の温度管理が重要な要素となる。
通常のビーカーや太い流路のある電気化学的な測定であれば液温の管理は容易であるが、本発明は密閉された微細流路中の反応を測定するカード型デバイスであって通常の温度管理は利用できない。 The diffusion coefficient D in the equation (1) is about 10 −9 m 2 / S in the case of an aqueous solution system, but the problem is that D has temperature dependence. This is because the diffusion coefficient varies depending on the viscosity of the liquid, and the viscosity varies depending on the temperature.
Therefore, when determining the concentration of the target substance by current, the temperature control of the liquid is an important factor.
Although the liquid temperature can be easily managed by an electrochemical measurement with a normal beaker or a thick channel, the present invention is a card type device for measuring a reaction in a sealed microchannel, Temperature management is not available.

そこで、第一の本発明は、前記カード型標的物質検出装置に、前記電極に移動する試料液の温度を検出するための温度検出器、及び前記電極に移動する試料液の温度を制御するための温度調節器を配置したものである。本発明のカードに、当該温度検出器と温度調節器を組み合せた例を図３に示す。   Therefore, the first aspect of the present invention provides the card-type target substance detection device with a temperature detector for detecting the temperature of the sample liquid moving to the electrode, and for controlling the temperature of the sample liquid moving to the electrode. The temperature controller is arranged. FIG. 3 shows an example in which the temperature detector and the temperature controller are combined with the card of the present invention.

図３の態様においては、温度調節器が、前記温度検出器により検出された温度に基づいてサーモモジュールを制御する温度制御部、及びサーモモジュールからなっている。ここで、温度検出器としては、熱電対や抵抗温度センサ等が用いられる。例えば、カード上の電極部（上部又は下部）又はコネクタ部に設けられた温度検出器により検出された温度変化は、コネクタを介して温度制御部に伝達される。サーモモジュールは、熱電半導体を利用したヒートポンプの一種であり、電流の方向を変更することにより冷却及び加熱のいずれかをすることができる装置であり、市販品を応用できる。電流の方向の変更は、温度検出器から伝達された温度変化に基づき、温度制御部で制御することにより容易に行うことができる。例えば、電極部の温度が３９℃を超えた場合には冷却し、３５℃未満の場合には加熱することにより、電極部の試料液の温度を３７±２℃に制御できる。   In the embodiment of FIG. 3, the temperature controller includes a temperature control unit that controls the thermo module based on the temperature detected by the temperature detector, and a thermo module. Here, a thermocouple, a resistance temperature sensor, or the like is used as the temperature detector. For example, a temperature change detected by a temperature detector provided in an electrode part (upper or lower part) or a connector part on the card is transmitted to the temperature control part via the connector. The thermo module is a kind of heat pump using a thermoelectric semiconductor, and is a device that can perform either cooling or heating by changing the direction of current, and a commercially available product can be applied. The change of the direction of the current can be easily performed by controlling the temperature control unit based on the temperature change transmitted from the temperature detector. For example, it is possible to control the temperature of the sample liquid in the electrode part to 37 ± 2 ° C. by cooling when the temperature of the electrode part exceeds 39 ° C. and heating when the temperature is lower than 35 ° C.

また、温度検出器は、前記のように熱電対や抵抗温度センサであってもよいが、作用電極又は対電極を、例えば図４のように配置することにより、温度検出器と兼用することもできる。この場合には、新たな温度検出器を必要とせず、特に好ましい。   In addition, the temperature detector may be a thermocouple or a resistance temperature sensor as described above, but the working electrode or the counter electrode may be arranged as shown in FIG. it can. In this case, a new temperature detector is not required, which is particularly preferable.

また、第２の本発明は、前記カード型標的物質検出装置に、前記電極に移動する試料液の温度を検出するための温度検出器、及び前記温度検出器により検出された温度に基づいて電気化学的に検出された測定値を補正するための測定値制御部を配置したものである。本発明のカードに、当該温度検出器と測定値制御部を組み合せた例を図５に示す。   Further, the second aspect of the present invention provides the card-type target substance detection device with a temperature detector for detecting the temperature of the sample liquid moving to the electrode, and an electric power based on the temperature detected by the temperature detector. A measurement value control unit for correcting a chemically detected measurement value is arranged. An example in which the temperature detector and the measurement value control unit are combined with the card of the present invention is shown in FIG.

第２の本発明における温度検出器は、前記第１の発明と同様のものがすべて使用できる。測定値制御部には、予め試料液の温度変化により電流値がどのように変化するかを測定することにより得られた補正値をインプットしておく。得られた電流値と試料液温度により、現在の測定値の正確な値からの誤差値を補正することができるように予めプログラムしておけばよい。このような補正値は、通常、標準物質を用いた測定値から予め求めておくことができる。   As the temperature detector in the second aspect of the present invention, all the same temperature detectors as in the first aspect of the present invention can be used. A correction value obtained by measuring in advance how the current value changes due to the temperature change of the sample solution is input to the measurement value control unit. What is necessary is just to program in advance so that the error value from the exact value of the present measured value can be corrected by the obtained current value and sample solution temperature. Such a correction value can usually be obtained in advance from a measured value using a standard substance.

このように測定値を自動的に補正することにより、試料の温度調節は必要なくなる。   By automatically correcting the measurement value in this way, it is not necessary to adjust the temperature of the sample.

図６には、第１及び第２の発明におけるカードと制御部とのコネクション形態の一例を示す。   FIG. 6 shows an example of a connection form between the card and the controller in the first and second inventions.

実施例１
図１、図２及び図３のカードを用い、図６のようにカード部と制御部をコネクションして実験を行った。試料としては、フェロセンを用い、緩衝液としてはマロネート緩衝液を用いた。温度調節を行うことなく、測定した結果を図７に示す。図７に示すように、試料液温度の変化に伴い、試料濃度が一定であるにもかかわらず得られた電流値が変化した。これにより、カード型デバイスでは、電流値が温度に依存して変化し、測定値の誤差を生じることがわかる。これに対し、温度調節を行うことにより、測定値の変化を制御することができた。 Example 1
Experiments were performed by using the cards of FIGS. 1, 2 and 3 and connecting the card unit and the control unit as shown in FIG. Ferrocene was used as a sample, and malonate buffer was used as a buffer. The measurement results are shown in FIG. 7 without adjusting the temperature. As shown in FIG. 7, with the change in the sample solution temperature, the obtained current value changed despite the sample concentration being constant. As a result, in the card type device, it can be seen that the current value changes depending on the temperature, resulting in an error in the measured value. On the other hand, the change in the measured value could be controlled by adjusting the temperature.

カード型標的物質検出装置の一例を示す概念図である。It is a conceptual diagram which shows an example of a card type target substance detection apparatus. 電極の配置の一例を示す（斜線部は、コネクタとの接触部を示す）。An example of the arrangement of the electrodes is shown (the hatched portion indicates the contact portion with the connector). 本発明カード型標的物質検出装置における、カード、温度検出器、温度制御及びサーモモジュールの組み合せの概念図を示す。The conceptual diagram of the combination of a card | curd, a temperature detector, temperature control, and a thermo module in the card | curd type target substance detection apparatus of this invention is shown. 作用電極を温度センサとする場合の電極の配置例を示す。An arrangement example of electrodes when the working electrode is a temperature sensor is shown. 本発明のカード型標的物質検出装置における、カード、温度検出器及び測定値制御部の組み合せの概念図を示す。The conceptual diagram of the combination of a card | curd, a temperature detector, and a measured value control part in the card | curd type target substance detection apparatus of this invention is shown. 本発明におけるカードと制御部とのコネクションの状態を示す。The state of the connection of the card | curd and control part in this invention is shown. 試料液の温度変化と電流値との関係を示す図である。It is a figure which shows the relationship between the temperature change of a sample liquid, and an electric current value.

Claims (7)

カード型基板上に、（１）１又は２以上の試料載置用凹部、及び（２）標的物質を電気化学的に検出するための電極を有し、当該（１）凹部及び（２）電極が毛細管力を有する流路で連結されてなるカード型標的物質検出装置において、前記電極に移動する試料液の温度を検出するための温度検出器、及び前記電極に移動する試料液の温度を制御するための温度調節器が配置されていることを特徴とするカード型標的物質検出装置。   On the card-type substrate, (1) one or two or more sample mounting recesses, and (2) an electrode for electrochemical detection of the target substance, the (1) recess and (2) electrode In a card-type target substance detection apparatus, wherein the temperature of the sample liquid moving to the electrode is detected, and the temperature of the sample liquid moving to the electrode is controlled A card-type target substance detection device, characterized in that a temperature controller is provided for the purpose. 前記温度調節器が、前記温度検出器により検出された温度に基づいてサーモモジュールを制御する制御部、及びサーモモジュールからなるものである請求項１記載のカード型標的物質検出装置。   The card-type target substance detection device according to claim 1, wherein the temperature controller includes a controller that controls a thermo module based on a temperature detected by the temperature detector, and a thermo module. 前記温度検出器が、カード上の電極部又はカード上の電極部と制御部とのコネクタ部に設置されているものである請求項１又は２記載のカード型標的物質検出装置。   The card type target substance detection device according to claim 1 or 2, wherein the temperature detector is installed in an electrode part on a card or a connector part between an electrode part on the card and a control part. 前記電極が、作用電極、対電極及び参照電極からなるものであり、前記温度検出器を、作用電極又は対電極が兼用するものである請求項１〜３のいずれか１項記載のカード型標的物質検出装置。   The card type target according to any one of claims 1 to 3, wherein the electrode comprises a working electrode, a counter electrode, and a reference electrode, and the temperature detector is also used by the working electrode or the counter electrode. Substance detection device. カード型基板上に、（１）１又は２以上の試料載置用凹部、及び（２）標的物質を電気化学的に検出するための電極を有し、当該（１）凹部及び（２）電極が毛細管力を有する流路で連結されてなるカード型標的物質検出装置において、前記電極に移動する試料液の温度を検出するための温度検出器、及び前記温度検出器により検出された温度に基づいて電気化学的に検出された測定値を補正するための測定値制御部が配置されていることを特徴とするカード型標的物質検出装置。   On the card-type substrate, (1) one or two or more sample mounting recesses, and (2) an electrode for electrochemical detection of the target substance, the (1) recess and (2) electrode In a card-type target substance detection device, wherein the card type target substance detection device is connected by a flow path having a capillary force, based on a temperature detector for detecting the temperature of the sample liquid moving to the electrode, and the temperature detected by the temperature detector A card type target substance detection device, wherein a measurement value control unit for correcting a measurement value detected electrochemically is arranged. 前記温度検出器が、カード上の電極部又はカード上の電極部と制御部とのコネクタ部に設置されているものである請求項５記載のカード型標的物質検出装置。   6. The card-type target substance detection device according to claim 5, wherein the temperature detector is installed in an electrode part on the card or a connector part between the electrode part on the card and the control part. 前記電極が、作用電極、対電極及び参照電極からなるものであり、前記温度検出器を、作用電極又は対電極が兼用するものである請求項５又は６記載のカード型標的物質検出装置。   The card type target substance detection device according to claim 5 or 6, wherein the electrode comprises a working electrode, a counter electrode, and a reference electrode, and the temperature detector is also used by the working electrode or the counter electrode.
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