JP2018036154A - Method for electrically detecting target material, method for determining amount of the material, detection system, amount determination system, and reagent - Google Patents

Method for electrically detecting target material, method for determining amount of the material, detection system, amount determination system, and reagent Download PDF

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JP2018036154A
JP2018036154A JP2016169810A JP2016169810A JP2018036154A JP 2018036154 A JP2018036154 A JP 2018036154A JP 2016169810 A JP2016169810 A JP 2016169810A JP 2016169810 A JP2016169810 A JP 2016169810A JP 2018036154 A JP2018036154 A JP 2018036154A
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target substance
substance
amount
compound
electrical change
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尭生 小野
Takao Ono
尭生 小野
松本　和彦
Kazuhiko Matsumoto
和彦 松本
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Osaka University NUC
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Abstract

PROBLEM TO BE SOLVED: To provide a method for detection a target material in a biochemical field, a method for determining the amount of the target material, means for detecting the target material, and means for determining the amount of the target material which can use a FET regardless of the size of the target material.SOLUTION: The method for electrically detecting a target material includes the steps of (1) binding a compound A which selectively binds with a target material and can generate a material with electric charges to the target material; (2) causing the compound A to generate the material with electric charges; and (3) detecting an electric change by the material with electric charges by a field-effect transistor.SELECTED DRAWING: Figure 1

Description

本発明は、生化学分野における標的物質の検出、定量方法、検出、定量手段及びその方法に使用される試薬に関する。   The present invention relates to detection, quantification methods, detection and quantification means of target substances in the biochemical field, and reagents used in the methods.

従来より、タンパク質、ウィルス、細菌等は、生化学の分野で標的物質として低容量で迅速な検出方法、手段が検討されている。これら標的物質の多くは表面に特異的な特性、（例えば、形状、タンパク質、表面電荷等）を有しており、この特異的な特性を利用した選択的な検出方法が提案されている。検出手段としては光学的手段、電界効果型トランジスタ（以下、ＦＥＴと略す）による電気的手段（非特許文献１）等が提案されているが、特にＦＥＴを利用した電気的手段は、高感度化が可能であり簡便なシステムが期待されている。   Conventionally, proteins, viruses, bacteria, and the like have been investigated as low-volume and rapid detection methods and means as target substances in the field of biochemistry. Many of these target substances have specific characteristics (for example, shape, protein, surface charge, etc.) on the surface, and selective detection methods using these specific characteristics have been proposed. As detection means, optical means, electric means using a field effect transistor (hereinafter abbreviated as FET) (Non-Patent Document 1), etc. have been proposed. Therefore, a simple system is expected.

大野等 ジャーナル オブ アメリカンケミカルソサイエティー １３２、ｐ１８０１２（２０１０）Ohno et al. Journal of American Chemical Society 132, p18012 (2010)

しかしながら、ＦＥＴでは生理的条件下での巨大な標的物質は検出が難しいという課題があった。つまり生理条件では塩濃度が高く、標的物質の表面電荷の変化を感知できるいわゆるデバイ長が１ｎｍ以下と狭い範囲でしかなく、生体内のタンパク質等の巨大分子の電荷を安定的かつ定量的に検出することは困難であった。   However, the FET has a problem that it is difficult to detect a huge target substance under physiological conditions. In other words, the salt concentration is high under physiological conditions, and the so-called Debye length that can sense changes in the surface charge of the target substance is only a narrow range of 1 nm or less, and the charge of macromolecules such as proteins in the living body can be detected stably and quantitatively It was difficult to do.

本発明は、サイズに関わらずＦＥＴを利用することができる生化学分野の標的物質の検出、定量方法、検出、定量手段を提供することを目的とする。   An object of the present invention is to provide a detection, quantification method, detection and quantification means for a target substance in the field of biochemistry that can utilize an FET regardless of its size.

本発明者らは、下記の発明によって課題を解決することを見出した。   The present inventors have found that the problems can be solved by the following invention.

（１） 試料中の標的物質の検出又は定量のために電気的変化を測定する方法であって、
１）標的物質に選択的に結合しかつ有電荷物質を発生可能な化合物Ａを、標的物質に結合させる工程、
２）化合物Ａに有電荷物質を発生させる工程、及び
３）所要容積中の有電荷物質による電気的変化を電界効果型トランジスタで測定する工 程、を有する方法。
（２） 前記所要容積が５００μｍ３以下である前記１記載の方法。
（３） 前記測定する工程では、有電荷物質の前記所要容積外への拡散を抑制する前記１または２に記載の方法。
（４） デバイスのデバイ長よりも標的物質の径が大きい前記１〜３いずれかに記載の方法。
（５） 前記試料中の標的物質の濃度が１μＭ以下である前記１〜４いずれかに記載の方法。
（６） 標的物質の検出又は定量のための電気的変化測定方法に使用される試薬であって、標的物質に選択的に結合しかつ有電荷物質を発生可能な化合物Ａを含む試薬。
（７） 前記１〜５いずれかに記載の方法で測定される電気的変化に関わるパラメータ量を、４）パラメータ量と標的物質量との作成済みの相関データに照会し、標的物質量を算出する工程を有する標的物質の定量方法。
（８） 前記パラメータ量は、前記有電荷物質発生後の経過時間に対する電気的変化の関数において、当初の接線傾きに対する所定比の接線傾きになるまでの経過時間である前記８記載の標的物質の定量方法。
（９） 標的物質の検出又は定量のために電気的変化を測定するシステムであって、
１）標的物質に選択的に結合しかつ有電荷物質を発生可能な化合物Ａを担持可能な反応ウェル、
２）反応ウェルにおいて化合物Ａが発生する有電荷物質による電気的変化を測定する電界効果型トランジスタ、および 前記電気的変化に関わるパラメータ量の情報を外部へ送信する送信手段、
を有するシステム。
（１０） 電気的変化に関わるパラメータ量の情報を受信する受信手段、
電気的変化に関わるパラメータ量と標的物質量との作成済み相関データが格納されたデータ格納手段、
前記受信手段の情報に基づくパラメータ量を、前記データ格納手段の前記相関データに照会し、標的物質量を算出する処理手段、および 前記標的物質量の情報を送信する送信手段、
を有する標的物質の定量システム。 (1) A method for measuring an electrical change for detection or quantification of a target substance in a sample,
1) A step of binding Compound A that selectively binds to a target substance and can generate a charged substance to the target substance,
2) a step of generating a charged substance in compound A; and 3) a step of measuring an electrical change caused by the charged substance in a required volume with a field effect transistor.
(2) The method according to 1 above, wherein the required volume is 500 μm 3 or less.
(3) The method according to 1 or 2, wherein, in the measuring step, the diffusion of the charged substance outside the required volume is suppressed.
(4) The method according to any one of 1 to 3, wherein the target substance has a diameter larger than the Debye length of the device.
(5) The method according to any one of 1 to 4 above, wherein the concentration of the target substance in the sample is 1 μM or less.
(6) A reagent for use in an electrical change measurement method for detection or quantification of a target substance, comprising a compound A that selectively binds to the target substance and can generate a charged substance.
(7) The parameter amount related to the electrical change measured by the method described in any one of 1 to 5 above is referred to 4) the created correlation data between the parameter amount and the target substance amount, and the target substance amount is calculated. A method for quantifying a target substance comprising the step of
(8) The parameter amount is an elapsed time until a tangential slope having a predetermined ratio with respect to an initial tangential slope in a function of an electrical change with respect to an elapsed time after generation of the charged substance. Quantitation method.
(9) A system for measuring an electrical change for detection or quantification of a target substance,
1) a reaction well capable of supporting compound A that can selectively bind to a target substance and generate a charged substance;
2) a field effect transistor for measuring an electrical change caused by a charged substance generated by the compound A in the reaction well, and a transmission means for transmitting information on the parameter amount related to the electrical change to the outside,
Having a system.
(10) Receiving means for receiving information on parameter quantities related to electrical changes;
A data storage means for storing the created correlation data between the parameter amount related to the electrical change and the target substance amount;
A parameter unit based on information of the receiving unit is referred to the correlation data of the data storage unit, a processing unit for calculating a target substance amount, and a transmitting unit for transmitting information on the target substance amount;
A target substance quantification system having

本発明によれば、塩濃度が高い生理条件下でも、生体内のタンパク質等の巨大な標的物質を検出することが可能となる。   According to the present invention, a huge target substance such as a protein in a living body can be detected even under physiological conditions with a high salt concentration.

本発明の実施態様の一例であるウレアーゼを用いたシステムを示す概略図である。It is the schematic which shows the system using the urease which is an example of the embodiment of this invention. 本発明の検出開始時及び終点時の電流変化を示す図である。It is a figure which shows the electric current change at the time of the detection start of this invention, and an end point.

以下、本発明の実施形態について述べる。
本発明では、生体内の低分子物質だけでなく、ウイルス、タンパク質等の巨大標的物質でも検出することを可能にするため、標的物質そのものを検出するのではなく、標的物質と選択的に結合し、そして所定の反応により有電荷物質を発生するような化合物Ａを利用する。つまり化合物Ａから発生した有電荷物質を検出することを特徴とする。 Hereinafter, embodiments of the present invention will be described.
In the present invention, it is possible to detect not only a low molecular substance in a living body but also a large target substance such as a virus or a protein. Therefore, instead of detecting the target substance itself, it selectively binds to the target substance. And a compound A that generates a charged substance by a predetermined reaction is used. That is, it is characterized by detecting a charged substance generated from the compound A.

本発明の実施態様の一例を、図１で説明する。標的物質１は、標的物質１と選択的に結合する部位４を有するＦＥＴにまず固定される。選択的に結合する部位３を修飾したウレアーゼ２を化合物Ａとし、その化合物Ａは、所定の尿素を導入した水系の反応場でアンモニアを発生させる。このアンモニアは、水中で有電荷物質であるアンモニウムイオンとなり、このアンモニウムイオンの発生がＦＥＴによって検出される。そして検出された電流量によって、アンモニウムイオンの定量が可能となる。標的物質１は、ＦＥＴ上に確実に補足することが容易である場合は、標的を選択的に補足する部位４を使用することなくＦＥＴに直接固定させることも可能である。   An example of an embodiment of the present invention will be described with reference to FIG. The target substance 1 is first fixed to the FET having the site 4 that selectively binds to the target substance 1. The urease 2 modified with the selectively binding site 3 is defined as compound A, and the compound A generates ammonia in an aqueous reaction field into which a predetermined urea is introduced. The ammonia becomes an ammonium ion which is a charged substance in water, and the generation of the ammonium ion is detected by the FET. Then, ammonium ions can be quantified based on the detected current amount. If it is easy to surely capture the target substance 1 on the FET, the target substance 1 can be directly fixed to the FET without using the portion 4 for selectively capturing the target.

アンモニウムイオンの発生量は、ウレアーゼと導入された尿素量によって支配されるため、あらかじめウレアーゼと尿素の反応速度を測定しておき、検出された電気量の所定時間での変化を追跡することで、選択的に結合したウレアーゼの量が求められる。この選択的に結合したウレアーゼの量は、標的物質の量と比例するため、結果として標的物質の定量が可能となる。部位４としては、抗体、糖鎖、アプタマー等を対象とすることができる。また本発明においては、標的物質そのものが化合物Ａの特徴と有する者も含まれる。例えばピロリ菌等自分自身がウレアーゼを有するものであってもよい。
本発明の標的物質は、生体等の生の試料中に存在する場合だけでなく、濃度既知の標準試料中の標的物質でもよい。この場合、標準試料と生の試料とをウェルに適用し、生の試料中の標的物質の濃度の大小に依存して電気的変化が増減する。 Since the generation amount of ammonium ions is governed by urease and the amount of urea introduced, by measuring the reaction rate of urease and urea in advance and tracking the change in the detected amount of electricity over a predetermined time, The amount of selectively bound urease is determined. Since the amount of this selectively bound urease is proportional to the amount of the target substance, the target substance can be quantified as a result. The site 4 can be an antibody, sugar chain, aptamer, or the like. In the present invention, the target substance itself includes those having the characteristics of Compound A. For example, H. pylori itself may have urease.
The target substance of the present invention may be a target substance in a standard sample having a known concentration as well as in a living sample such as a living body. In this case, the standard sample and the raw sample are applied to the well, and the electrical change increases or decreases depending on the concentration of the target substance in the raw sample.

＜電気的検出方法＞
本発明の標的物質の電気的検出方法は、標的物質の検出方法であって、
１）標的物質に選択的に結合し、かつ有電荷物質を発生可能な化合物Ａを、標的物質に結合させる工程
２）化合物Ａに有電荷物質を発生させる工程
３）有電荷物質による電気的変化を電界効果型トランジスタで測定する工程
を有する。 <Electrical detection method>
The target substance electrical detection method of the present invention is a target substance detection method,
1) A step of binding compound A that selectively binds to a target substance and can generate a charged substance to the target substance 2) A step of generating a charged substance in compound A 3) An electrical change by the charged substance Is measured by a field effect transistor.

本発明では、試料中の標的物質を選択的にＦＥＴ上に固定するために、ＦＥＴ上には標的物質を選択的に固定するための捕捉物質層が設けられている。これは常法により作製することができる捕捉層である。   In the present invention, in order to selectively fix the target substance in the sample on the FET, a capture substance layer for selectively fixing the target substance is provided on the FET. This is a trapping layer that can be made by conventional methods.

ついで、標的物質に選択的に結合し、かつ有電荷物質を発生可能な化合物Ａを、標的物質に結合させる。本発明の標的物質に選択的に結合し、かつ有電荷物質を発生可能な化合物Ａとは、係る２つの機能を有する物質であれば特に限定はされないが、酵素として機能する部分と標的物質と選択的に結合する部分を有する物質が好ましい。例えばウレアーゼを修飾した抗体等が挙げられる。この化合物Ａは単独の試薬として利用することができる。   Next, the compound A that selectively binds to the target substance and can generate a charged substance is bound to the target substance. The compound A that selectively binds to the target substance of the present invention and can generate a charged substance is not particularly limited as long as it is a substance having such two functions. Substances having moieties that bind selectively are preferred. Examples thereof include antibodies modified with urease. This compound A can be used as a single reagent.

有電荷物質とは、ＦＥＴで検出することができる電荷を有する物質を意味し、ＦＥＴのデバイ長が１ｎｍ以下の範囲で測定可能な分子サイズを有するものが好ましい。例えばアンモニアが挙げられる。   The charged substance means a substance having a charge that can be detected by the FET, and preferably has a molecular size that can be measured when the Debye length of the FET is 1 nm or less. An example is ammonia.

本発明では、まず標的物質を前記化合物Ａと結合させる工程を有する。ついで、化合物Ａに有電荷物質を発生させる工程を有するが、この工程では、化合物Ａ自身が有電荷物質を発生させてもよいし、前述の尿素のような助化合物を使用し、有電荷物質を発生させてもよい。用途によって適宜選択の幅が拡大することができる点で、助化合物を使用する方法が好ましい。   In the present invention, first, a target substance is bound to the compound A. Next, the compound A has a step of generating a charged substance. In this step, the compound A itself may generate a charged substance, or the above-mentioned auxiliary compound such as urea is used to charge the charged substance. May be generated. A method using an auxiliary compound is preferable in that the range of selection can be appropriately expanded depending on the application.

本発明では、有電荷物質を電気的検出手段により検出するが、電気的検出方法は感度の点からＦＥＴによるものが好ましく、特にチャネル部がグラフェン、シリコン、カーボンナノチューブをはじめとする半導体、金属、酸化物を使用したものが好ましい。   In the present invention, a charged substance is detected by an electrical detection means, and the electrical detection method is preferably based on FETs from the viewpoint of sensitivity. In particular, the channel portion is a semiconductor including graphene, silicon, carbon nanotube, metal, Those using oxides are preferred.

本発明では、化合物Ａに有電荷物質を発生させる工程、検出する工程の容積が５００μｍ３以下であることが好ましい。反応場を限定することにより感度の向上を図ることができる。例えば高さ４μｍ、幅１０μｍ、奥行き１０μｍのウェル内で検出する方法を実施することが好ましい。高さは、デバイ長以上であることから１ｎｍ以上あることが好ましい。検出する工程の容積としては１ピコリットル〜１フェムトリットルが好ましい。ウェルは、１層、２層、或いは多層の材料からなる。例えば２層の場合、アルミナとフッ素樹脂の積層材料が挙げられる。 In the present invention, the volume of the step of generating a charged substance in compound A and the step of detecting is preferably 500 μm 3 or less. The sensitivity can be improved by limiting the reaction field. For example, it is preferable to carry out a detection method in a well having a height of 4 μm, a width of 10 μm, and a depth of 10 μm. The height is preferably 1 nm or more because it is the Debye length or more. The volume of the detecting step is preferably 1 picoliter to 1 femtoliter. The well is made of one, two, or multiple layers of material. For example, in the case of two layers, a laminate material of alumina and a fluororesin can be used.

また有電荷物質が揮発性の高い物質であったり、容易に外気の影響を受けやすい物質である場合は、前記検出する工程では、有電荷物質を前記容積に保持する手段が設けられていることが好ましい。この手段の存在により検出精度が安定する。手段としては、特に限定はないが、樹脂性の蓋、空気層が挙げられる。   In addition, when the charged substance is a highly volatile substance or a substance that is easily affected by outside air, a means for holding the charged substance in the volume is provided in the detection step. Is preferred. The presence of this means stabilizes the detection accuracy. The means is not particularly limited, and examples thereof include a resinous lid and an air layer.

本発明では、生体内の低分子物質だけでなく、ウイルス、タンパク質等の巨大標的物質でも検出することを可能にする。したがって、デバイスのデバイ長よりも標的物質の径が大きい場合、本発明の効果が期待される。   In the present invention, it is possible to detect not only a low molecular substance in a living body but also a huge target substance such as a virus or a protein. Therefore, when the diameter of the target substance is larger than the Debye length of the device, the effect of the present invention is expected.

ここで本発明が標的物質とするウイルス、タンパク質等は、それぞれ固有の形状を有していることから一義的に標的物質の径を定めることができないが、本発明では検出可能なデバイ長よりも標的物質が大きく、そのデバイ長では安定的・定量的な検出ができない大きさを意味する。
本発明では高感度の検出が可能であり、標的物質の濃度が１μＭ以下であることが可能である。 Here, viruses, proteins, and the like that are targeted by the present invention have unique shapes, and thus the diameter of the target substance cannot be uniquely determined. This means that the target substance is large and cannot be detected stably and quantitatively by its Debye length.
In the present invention, highly sensitive detection is possible, and the concentration of the target substance can be 1 μM or less.

＜定量方法＞
本発明では、電気的変化に関わるパラメータ量と標的物質量との作成済みの相関データとを比較することにより標的物質を定量することが可能である。
例えば、前述のウレアーゼと尿素の系では、発生するアンモニウムイオン量による電気的変化に関わるパラメータ（ここでは電流値）とウレアーゼ量の検量線を作成しておき、その検量線からウレアーゼ量を知ることができ、結果として標的物質の定量が可能となる。
検量線としては、他に作成しているデータベース上のデータと比較対応使用することも可能である。 <Quantitative method>
In the present invention, the target substance can be quantified by comparing the parameter amount related to the electrical change and the created correlation data between the target substance amount.
For example, in the urease-urea system described above, create a calibration curve for the parameters (current value in this case) and the amount of urease related to the electrical change due to the amount of ammonium ions generated, and know the amount of urease from the calibration curve. As a result, the target substance can be quantified.
As a calibration curve, it is also possible to use it for comparison with data on other databases created.

本発明での検出速度は、例えば前述の系では、ウレアーゼと導入された尿素量によって支配されるため、あらかじめウレアーゼと尿素の反応速度を測定しておき、検出された電気量の所定時間での変化を追跡することで、選択的に結合したウレアーゼの量が求められる。   For example, in the above-described system, the detection speed in the present invention is governed by the amount of urease and urea introduced, so the reaction rate of urease and urea is measured in advance and the detected amount of electricity at a predetermined time is measured. By tracking changes, the amount of selectively bound urease is determined.

この場合電気変化量は、当初の接線の傾きに対する所定値の傾きになるまでの時間に基づき判断される。このことは、ＦＥＴの感度の良し悪しに関係なく検出、定量することができることを意味し、検出、定量のデータバラツキを小さくすることが可能となる。   In this case, the electrical change amount is determined based on the time until the inclination of the predetermined value with respect to the initial inclination of the tangent line. This means that detection and quantification can be performed regardless of whether the sensitivity of the FET is good or bad, and data variation in detection and quantification can be reduced.

＜システム＞
本発明では、標的物質の検出又は定量のために電気的変化を測定するシステムであって、１）標的物質に選択的に結合しかつ有電荷物質を発生可能な化合物Ａを担持可能な反応ウェル、２）反応ウェルにおいて化合物Ａが発生する有電荷物質による電気的変化を測定する電界効果型トランジスタ、および、前記電気的変化に関わるパラメータ量の情報を外部へ送信する送信手段、を有するシステムを構成する。 <System>
The present invention is a system for measuring an electrical change for detection or quantification of a target substance, and 1) a reaction well capable of carrying compound A that can selectively bind to the target substance and generate a charged substance. 2) a system having a field-effect transistor that measures an electrical change caused by a charged substance generated by compound A in a reaction well, and a transmission unit that transmits information on parameter amounts related to the electrical change to the outside. Configure.

そして、電気的変化に関わるパラメータ量の情報を受信する受信手段、電気的変化に関わるパラメータ量と標的物質量との作成済み相関データが格納されたデータ格納手段、前記受信手段の情報に基づくパラメータ量を、前記データ格納手段の前記相関データに照会し、標的物質量を算出する処理手段、および前記標的物質量の情報を送信する送信手段、を有する標的物質の定量システムを構成することが可能である。
本発明の手段は、それぞれ独立した手段であって、全体としてシステムを構成することができる。 And receiving means for receiving information on the parameter amount relating to the electrical change, data storing means for storing the created correlation data between the parameter quantity relating to the electrical change and the target substance amount, and parameters based on the information of the receiving means It is possible to configure a target substance quantification system having processing means for referring to the correlation data of the data storage means, calculating target substance quantity, and transmitting means for transmitting information on the target substance quantity. It is.
The means of the present invention are independent means, and can constitute a system as a whole.

１ 標的物質
２ ウレアーゼ
３ 標的と選択的に結合する部位（検出抗体）
４ 標的を選択的に補足する部位（捕捉抗体）
５ 容積を保持する手段
Ｇ グラフェン（チャネル）
Ｓ ソース電極
Ｄ ドレイン電極
ＳｉＯ２ 基体
１０ 反応開始当初の電流変化を示す接線
１１ 反応終点の電流変化を示す接線 1 Target substance 2 Urease 3 Site that selectively binds to target (detection antibody)
4 Site that selectively captures target (capture antibody)
5 Means to hold volume G Graphene (channel)
S source electrode D drain electrode SiO2 substrate 10 tangent indicating current change at the beginning of reaction 11 tangent indicating current change at the end of reaction

Claims (10)

試料中の標的物質の検出又は定量のために電気的変化を測定する方法であって、
１）標的物質に選択的に結合しかつ有電荷物質を発生可能な化合物Ａを、標的物質に結合させる工程、
２）化合物Ａに有電荷物質を発生させる工程、及び
３）所要容積中の有電荷物質による電気的変化を電界効果型トランジスタで測定する工程、を有する方法。 A method for measuring an electrical change for detection or quantification of a target substance in a sample, comprising:
1) A step of binding Compound A that selectively binds to a target substance and can generate a charged substance to the target substance,
2) a step of generating a charged substance in the compound A, and 3) a step of measuring an electrical change caused by the charged substance in a required volume with a field effect transistor. 前記所要容積が５００μｍ３以下である請求項１記載の方法。 The method according to claim 1, wherein the required volume is 500 μm 3 or less. 前記測定する工程では、有電荷物質の前記所要容積外への拡散を抑制する請求項１または２に記載の方法。   The method according to claim 1, wherein in the measuring step, diffusion of the charged substance out of the required volume is suppressed. デバイスのデバイ長よりも標的物質の径が大きい請求項１〜３いずれかに記載の方法。   The method according to claim 1, wherein the diameter of the target substance is larger than the Debye length of the device. 前記試料中の標的物質の濃度が１μＭ以下である請求項１〜４いずれかに記載の方法。   The method according to claim 1, wherein the concentration of the target substance in the sample is 1 μM or less. 標的物質の検出又は定量のための電気的変化測定方法に使用される試薬であって、標的物質に選択的に結合しかつ有電荷物質を発生可能な化合物Ａを含む試薬。   A reagent used for an electrical change measurement method for detection or quantification of a target substance, comprising a compound A that selectively binds to a target substance and can generate a charged substance. 請求項１〜５いずれかに記載の方法で測定される電気的変化に関わるパラメータ量を、４）パラメータ量と標的物質量との作成済みの相関データに照会し、標的物質量を算出する工程、を有する標的物質の定量方法。   A step of calculating the target substance amount by referring to the already created correlation data between the parameter quantity and the target substance quantity for the parameter quantity relating to the electrical change measured by the method according to claim 1. A method for quantifying a target substance having 前記パラメータ量は、前記有電荷物質発生後の経過時間に対する電気的変化の関数において、当初の接線傾きに対する所定比の接線傾きになるまでの経過時間である請求項7載の標的物質の定量方法。   8. The method for quantifying a target substance according to claim 7, wherein the parameter amount is an elapsed time until a tangential slope having a predetermined ratio with respect to an initial tangential slope in a function of an electrical change with respect to the elapsed time after generation of the charged substance. . 標的物質の検出又は定量のために電気的変化を測定するシステムであって、
１）標的物質に選択的に結合しかつ有電荷物質を発生可能な化合物Ａを担持可能な反応ウェル、
２）反応ウェルにおいて化合物Ａが発生する有電荷物質による電気的変化を測定する電界効果型トランジスタ、および 前記電気的変化に関わるパラメータ量の情報を外部へ送信する送信手段、
を有するシステム。 A system for measuring electrical changes for detection or quantification of a target substance,
1) a reaction well capable of supporting compound A that can selectively bind to a target substance and generate a charged substance;
2) a field effect transistor for measuring an electrical change caused by a charged substance generated by the compound A in the reaction well, and a transmission means for transmitting information on the parameter amount related to the electrical change to the outside,
Having a system. 電気的変化に関わるパラメータ量の情報を受信する受信手段、
電気的変化に関わるパラメータ量と標的物質量との作成済み相関データが格納されたデータ格納手段、
前記受信手段の情報に基づくパラメータ量を、前記データ格納手段の前記相関データに照会し、標的物質量を算出する処理手段、および
前記標的物質量の情報を送信する送信手段、
を有する標的物質の定量システム。 Receiving means for receiving information on parameter quantities related to electrical changes,
A data storage means for storing the created correlation data between the parameter amount related to the electrical change and the target substance amount;
Processing means for calculating the target substance amount by referring to the correlation data of the data storage means for the parameter amount based on the information of the receiving means, and transmitting means for transmitting the target substance amount information;
A target substance quantification system having
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