JPH0552836A - Displaying method for completion of test and detection tester fitted with test completion display - Google Patents
Displaying method for completion of test and detection tester fitted with test completion displayInfo
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- JPH0552836A JPH0552836A JP24021791A JP24021791A JPH0552836A JP H0552836 A JPH0552836 A JP H0552836A JP 24021791 A JP24021791 A JP 24021791A JP 24021791 A JP24021791 A JP 24021791A JP H0552836 A JPH0552836 A JP H0552836A
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Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、クロマトグラフ媒体等
の毛管移動媒体を使用して被検査流体中の特定物質を検
出試験する方法および装置であって、特に試験の終了を
呈色表示するようにした方法および装置に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and apparatus for detecting and testing a specific substance in a fluid to be inspected by using a capillary moving medium such as a chromatographic medium, and in particular, indicates the end of the test with a color. Method and apparatus.
【０００２】[0002]
【従来の技術】クロマトグラフ法等により被検査流体中
の特定物質を検出試験する際に、試験終了の表示が必要
な場合がある。従来は、クロマトグラフ媒体の下流終端
付近にｐＨ指示薬や水分試験紙を施して、試験の終了を
色の変化により表示または検知している。ｐＨ指示薬に
よる方法は、被検査流体がｐＨ指示薬を施した下流終端
位置まで毛管移動した時のｐＨ変化に伴う呈色状態の変
化を利用したものである。また、水分試験紙による方法
は、被検査液体が毛管移動によって水分試験紙を配置し
た下流終端位置に到達した際の、試験紙の液体吸収に伴
う色の変化を利用したものである。2. Description of the Related Art When a test for detecting a specific substance in a fluid to be inspected is carried out by a chromatographic method or the like, it is sometimes necessary to indicate the end of the test. Conventionally, a pH indicator or a moisture test paper is applied near the downstream end of the chromatographic medium, and the end of the test is displayed or detected by a color change. The method using a pH indicator utilizes the change in the coloration state accompanying the pH change when the test fluid moves to the downstream end position where the pH indicator is applied to the downstream end position. Further, the method using a moisture test paper utilizes a change in color due to liquid absorption of the test paper when the liquid to be inspected reaches the downstream end position where the moisture test paper is arranged by capillary movement.
【０００３】しかしながら、ｐＨ指示薬は通常最初から
一定の色を有しており、無色から有色に変化するものは
極めて少ない。しかも、ｐＨの変化前の色と変化後の色
とが近似している場合が多く、試験の終了を明確に表示
しにくいという欠点がある。更に、ｐＨ指示薬のある呈
色表示部の前位置等に、ｐＨを安定させるための緩衝液
域を設けておく必要がある。However, pH indicators usually have a constant color from the beginning, and very few change from colorless to colored. In addition, the color before the change of the pH and the color after the change of the pH are often similar to each other, and there is a drawback that it is difficult to clearly indicate the end of the test. Further, it is necessary to provide a buffer solution region for stabilizing the pH at a position in front of the color display unit having a pH indicator.
【０００４】また、水分試験紙の場合は、紙を着色して
おかないと、乾燥状態と吸水状態とで色の違いを生じな
いため、やはり最初から一定の色に着色されており、し
かも乾燥時の色と吸水時の色は同系色であるために、色
の区別がしにくく、試験の終了を明確に表示できない。
その上、水分試験紙位置まで到達した被検査液体がその
後の時間経過に伴って蒸発してしまうと水分試験紙が再
度乾燥して、元の色に戻ってしまい、試験の終了を表示
できなくなるという欠点がある。Further, in the case of the moisture test paper, unless the paper is colored, there is no difference in color between the dry state and the water absorbing state. Therefore, the moisture test paper is also colored in a constant color from the beginning and is dried. Since the color at the time and the color at the time of absorbing water are similar colors, it is difficult to distinguish the colors and the end of the test cannot be clearly displayed.
In addition, if the liquid to be inspected that has reached the position of the moisture test paper evaporates with the passage of time thereafter, the moisture test paper dries again and returns to the original color, making it impossible to display the end of the test. There is a drawback.
【０００５】[0005]
【発明の内容】本発明者らは上記した従来技術における
ような欠点のない、毛管移動媒体による検出試験の終了
を明確に表示できる方法を求めて研究を行ってきた。そ
の結果、クロマトグラフ媒体等の毛管移動媒体の所定位
置に有色体を施す共に、毛管移動媒体の下流終端または
その付近に有色体用の捕捉材を位置させ、被検査流体の
毛管移動に伴って下流終端部へと移動してきた有色体を
該捕捉材で捕捉させると、その部分に鮮明な呈色が発現
して試験の終了を明確に表示できることを見出した。The present inventors have conducted research for a method capable of clearly indicating the end of a detection test using a capillary moving medium, which does not have the above-mentioned drawbacks of the prior art. As a result, the colored body is applied to a predetermined position of the capillary moving medium such as a chromatographic medium, and the capturing material for the colored body is positioned at or near the downstream end of the capillary moving medium, so that the fluid to be inspected moves along the capillary. It has been found that when the colored material that has moved to the downstream end portion is captured by the capturing material, a clear color is developed in that portion and the end of the test can be clearly displayed.
【０００６】更に、本発明者らは、上記の有色体および
その捕捉材の組み合わせの代わりに、反応型発色剤を使
用して、毛管移動媒体の所定位置に反応型発色剤の１つ
または２つ以上の成分を施すと共に、下流終端またはそ
の付近に反応型発色剤の残りの成分を位置させ、被検査
流体の毛管移動に伴って下流終端へと移動してきた反応
型発色剤の該１つまたは２つ以上の成分を該残りの成分
と反応させ発色させた場合にも、同様に鮮明な発色また
は色変化を生じて試験の終了を明確に表示できることを
見出した。Furthermore, the present inventors have used a reactive color former instead of the combination of the above-mentioned colored body and its scavenger, and used one or two of the reactive color formers at a predetermined position of the capillary transfer medium. One or more components are applied, and the remaining components of the reactive color former are located at or near the downstream end, and the one of the reactive color formers that has moved to the downstream end with the capillary movement of the fluid to be tested. It has also been found that even when two or more components are reacted with the remaining components to cause color development, similarly vivid color development or color change occurs and the end of the test can be clearly displayed.
【０００７】したがって、本発明は、毛管移動媒体を
使用して被検査流体中の特定物質を検出試験する方法に
おいて、毛管移動媒体の所定位置に有色体を施すと共
に、毛管移動媒体の下流終端またはその付近に有色体用
の捕捉材を位置させ、被検査流体の毛管移動に伴って下
流終端側に移動してきた有色体を捕捉材で捕捉して試験
の終了を呈色表示することを特徴とする方法である。Therefore, the present invention provides a method for detecting and testing a specific substance in a fluid to be inspected by using a capillary moving medium, in which a colored body is applied to a predetermined position of the capillary moving medium and a downstream end of the capillary moving medium or A capturing material for a colored body is located in the vicinity of the colored body, and the colored body that has moved to the downstream end side along with the capillary movement of the fluid to be inspected is captured by the capturing material and the end of the test is displayed in color. Is the way to do it.
【０００８】更に、本発明は、毛管移動媒体を使用し
て被検査流体中の特定物質を検出試験する方法におい
て、毛管移動媒体の所定位置に反応型発色剤の１つまた
は２つ以上の成分を施すと共に、毛管移動媒体の下流終
端またはその付近に反応型発色剤の残りの成分を位置さ
せ、被検査流体の毛管移動に伴って下流終端側に移動し
てきた反応型発色剤の該１つまたは２つ以上の成分を該
残りの成分と反応・発色させて試験の終了を呈色表示す
ることを特徴とする方法である。また、上記の方法と
の方法は併用してもよく、したがって本発明は、上記
の方法との方法を組み合わせた方法をも包含する。Furthermore, the present invention provides a method for detecting and testing a specific substance in a fluid to be inspected using a capillary transfer medium, wherein one or more components of a reactive color former are present at a predetermined position of the capillary transfer medium. And the remaining components of the reactive color former are positioned at or near the downstream end of the capillary transfer medium, and the one of the reactive color formers that has moved to the downstream end side with the capillary movement of the fluid to be tested. Alternatively, the method is characterized in that two or more components are reacted and colored with the remaining components to color-code the end of the test. Further, the method and the method described above may be used in combination, and thus the present invention also includes a method in which the method and the method described above are combined.
【０００９】更に、本発明は上記した方法を実施するの
に使用する、毛管移動媒体、その所定位置に施した有色
体および／或いは反応型発色剤の１つまたは２つ以上の
成分、並びに毛管移動媒体の下流終端またはその付近に
配置した有色体用の捕捉材および／または反応型発色剤
の残りの成分を備えていることを特徴とする試験終了表
示体付きの検出試験装置を包含する。Furthermore, the present invention is used to carry out the above-described method, a capillary transfer medium, one or more components of a colored body and / or a reactive color former applied in its position, and a capillary. A detection test device with an end-of-test indicator, characterized in that the detection test device is provided with a scavenger for a colored body and / or the remaining components of a reactive color former disposed at or near the downstream end of the moving medium.
【００１０】上記において「毛管移動媒体を使用した検
出試験」とは、毛管媒体中を被検査流体を毛管移動させ
てその中に含まれる各成分の分配係数や吸着程度の差を
利用して特定の物質を検出したり、分離する検出試験を
いい、その代表例は毛管移動媒体として紙、布帛、粒状
物等のクロマトグラフ媒体を用いたクロマトグラフ法を
挙げることができる。しかしながら、クロマトグラフ法
に限定されず、毛管移動媒体を利用する同種の検出試験
法は、いずれも本発明に包含される。また、上記におい
て「毛管移動媒体の下流終端」とは、毛管移動媒体の試
料採取部を設ける一方の側を上流側とし、被検査流体が
該上流側から毛管移動媒体の他方の端部に向かって毛管
移動していく該他方の側を下流側とし、下流終端とは毛
管移動媒体の該下流側の端部をいう。更に「下流終端の
付近」とは、毛管移動媒体の下流終端の近傍であって、
下流終端に到達する前の位置をいう。In the above description, the "detection test using a capillary moving medium" is specified by moving the fluid to be inspected in the capillary medium by capillarity and utilizing the difference in distribution coefficient and adsorption degree of each component contained therein. Is a detection test for detecting or separating the substance, and a typical example thereof is a chromatographic method using a chromatographic medium such as paper, cloth, or granular material as a capillary moving medium. However, the present invention is not limited to the chromatographic method, and any detection test method of the same type using a capillary transfer medium is included in the present invention. Further, in the above description, “downstream end of the capillary moving medium” refers to one side where the sampling portion of the capillary moving medium is provided as the upstream side, and the fluid to be tested goes from the upstream side to the other end of the capillary moving medium. The other side of the capillary moving medium is the downstream side, and the downstream end means the downstream end of the capillary moving medium. Furthermore, "near the downstream end" is near the downstream end of the capillary moving medium,
The position before reaching the downstream end.
【００１１】上記の方法で使用する有色体とは、それ
自体が毛管移動媒体や被検査流体とは異なる色を有し、
捕捉材を配置した位置で捕捉材により捕捉されて有色体
自体の色を発現し、試験の終了を表示し得る着色体をい
う。有色体と捕捉材との組み合わせとしては、有色体と
捕捉材との間の物理的な付加作用や吸着作用を利用する
もの、化学反応や生物学的反応を伴って有色体を捕捉材
位置に捕捉すもの等のいずれをも採用できる。そのうち
でも、有色体として有色磁性粒子等の有色磁性体を使用
し、捕捉材として磁石等の磁場発生部材を使用して、該
有色磁性体を磁力によって毛管移動媒体の下流終端また
はその付近に捕捉して有色体の色に基づく呈色を生じさ
せるのが、簡単で便利である。The colored body used in the above method has a color different from that of the capillary moving medium or the fluid to be inspected,
A colored body that can be captured by the capturing material at the position where the capturing material is arranged to develop the color of the colored body itself and indicate the end of the test. As the combination of the colored body and the capturing material, those utilizing physical addition or adsorption between the colored body and the capturing material, the colored body to the capturing material position along with a chemical reaction or a biological reaction. Any one such as a capturing device can be adopted. Among them, a colored magnetic material such as colored magnetic particles is used as the colored material, and a magnetic field generation member such as a magnet is used as the capturing material, and the colored magnetic material is captured by magnetic force at the downstream end of the capillary moving medium or in the vicinity thereof. It is easy and convenient to generate a coloration based on the color of the colored body.
【００１２】また、上記の方法で使用する反応型発色
剤とは、２または３以上の反応性成分からなっていて、
それらが反応することによって無色から特定の色に発色
するか、または最初の色とは異なった色に色変化する発
色剤をいう。反応型発色剤としては、化学反応を伴う発
色剤、生物反応を伴う発色剤、またはその両方を伴う発
色剤のいずれをも使用することができる。例えば、検出
試験装置が、イムノアッセイ等に利用されるものの場合
は、反応型発色剤の１つまたは２つ以上の成分を酵素発
色性基質または酵素として、それを毛管移動媒体の所定
位置に施し、残りの成分をそれに対応した酵素または酵
素発色性基質として、それを毛管移動媒体の下流終端ま
たはその付近に位置させて、両者の反応による発色また
は色変化により試験の終了を表示するようにするとよ
い。The reactive color former used in the above method is composed of 2 or 3 or more reactive components,
A color-forming agent that reacts with them to develop a color from a colorless color to a specific color or to change a color different from the initial color. As the reactive color former, a color former involving a chemical reaction, a color former involving a biological reaction, or a color former involving both of them can be used. For example, when the detection test device is used for an immunoassay or the like, one or more components of the reactive chromogenic agent are used as enzyme chromogenic substrates or enzymes, and they are applied to predetermined positions of the capillary migration medium, It is advisable to use the remaining components as the corresponding enzymes or enzyme color-developing substrates, and locate them at or near the downstream end of the capillary migration medium, so that the color development or color change resulting from the reaction of both indicates the end of the test. ..
【００１３】そして、上記で使用する有色体およびその
捕捉材並びに反応型発色剤としては、各々の検出試験の
種類や内容に応じて、実施しようとしている検出試験の
妨害にならないものを選択して使用するのがよい。ま
た、有色体および反応型発色剤の該１つまたは２つ以上
の成分は、途中で毛管移動媒体に吸着されたり付着する
ことがなく、被検査流体の毛管移動に伴って毛管移動媒
体の下流終端まで円滑に移動できるものを選択して使用
するのがよい。また、有色体およびその捕捉材並びに反
応型発色剤の該１つまたは２つ以上の成分と該残りの成
分は、毛管移動媒体上の検出試験の妨げにならず、しか
も試験の終了を明確に表示し得る位置にそれぞれ設ける
必要がある。As the colored body, the scavenger therefor and the reactive color former used above, those which do not interfere with the detection test to be carried out are selected according to the type and content of each detection test. Good to use. Further, the one or more components of the colored body and the reactive color former are not adsorbed or adhered to the capillary moving medium on the way, and are downstream of the capillary moving medium as the test fluid moves by the capillary. It is better to select and use one that can move smoothly to the end. In addition, the one or more components and the remaining components of the colored body and its scavenger and the reactive color former do not interfere with the detection test on the capillary migration medium, and clearly indicate the end of the test. It must be provided at each position where it can be displayed.
【００１４】通常、捕捉材および／または反応型発色剤
の該残りの成分は、毛管移動媒体上の下流終端またはそ
の付近に適当な手段により固定して設けるのがよい。ま
た、有色体および／或いは反応型発色剤の該１つまたは
２つ以上の成分は、捕捉材および／または反応型発色剤
の該残りの成分を設けた位置から所定の間隔をあけて、
それよりも上流側に、被検査流体の毛管移動に伴って下
流側に移動し得るようにして施すのがよい。その間隔
は、検出試験装置の未使用時に両者間で捕捉や反応が生
じないような距離としておく。有色体および反応型発色
剤の該１つまたは２つ以上の成分が検出試験を何ら妨害
しない場合には、それらを毛管移動媒体の上流端側に施
してもよいが、通常は、検出しようとする特定物質の検
出域から外れた下流側に設けるのが安全であり好まし
い。Usually, the remaining components of the scavenger and / or the reactive color former are preferably provided by a suitable means at or near the downstream end of the capillary transfer medium. Further, the one or more components of the colored body and / or the reactive color former are separated from the position at which the scavenger and / or the remaining component of the reactive color former are provided,
It is advisable to perform the treatment on the upstream side so that the fluid to be inspected can move to the downstream side as the capillary moves. The interval is set such that no capture or reaction occurs between the two when the detection test device is not used. If the colored body and the one or more components of the reactive color former do not interfere with the detection test in any way, they may be applied to the upstream end of the capillary transfer medium, but it is usually not It is safe and preferable to provide it on the downstream side outside the detection range of the specified substance.
【００１５】また、毛管移動媒体の下流終端またはその
付近に捕捉材および／または反応型発色剤の該残りの成
分を配置するに当たって、その配置形状は毛管移動媒体
を横切る直線状としたり、花型、星型、その他の任意の
形状にすることができ、その場合にはその配置形状に応
じた直線状、花型、星型等の呈色状態を発現させること
ができ、試験の終了を一層明確に表示することができ
る。In arranging the remaining components of the capturing material and / or the reactive color former at or near the downstream end of the capillary moving medium, the arrangement shape may be a straight line crossing the capillary moving medium, or a flower shape. , Star shape, or any other shape, and in that case, it is possible to develop a linear, flower-shaped, star-shaped coloration state according to the arrangement shape, and to further complete the test. Can be clearly displayed.
【００１６】本発明の内容の理解のために、近年開発さ
れたイムノクロマトグラフ法（U.R.Charaborty et al.,
Ann. Biol. clin.,1990,48, 403-408 ; R.F. Zuk et a
l.,CLINICAL CHEMISTRY, Vol.31, No.7, 1985）を例に
とって、本発明を図１〜２により具体的に説明する。In order to understand the content of the present invention, an immunochromatographic method recently developed (URCharaborty et al.,
Ann. Biol. Clin., 1990,48, 403-408; RF Zuk et a
1, CLINICAL CHEMISTRY, Vol. 31, No. 7, 1985) as an example, the present invention will be described in detail with reference to FIGS.
【００１７】図１および図２において、１はクロマトグ
ラフ媒体、２は裏打ちフイルム、３は試料（検体）採取
部、４は検体中の測定対象抗原と特異的に反応する標識
した抗体（以後「標識抗体」という）の乾燥物を施した
位置、５は検体中の測定対象抗原と別の位置で特異的に
反応する抗体（以後「固定化抗体」という）をクロマト
グラフ媒体に固定化してある位置（検出部）、６は有色
磁性粒子、７は磁石（ゴム磁石等）、８は酵素発色性基
質（酵素）、そして９は固定化された酵素（酵素発色性
基質）を示す。In FIGS. 1 and 2, 1 is a chromatographic medium, 2 is a backing film, 3 is a sample (specimen) sampling portion, 4 is a labeled antibody that specifically reacts with an antigen to be measured in the specimen (hereinafter referred to as " A dried product of "labeled antibody") is provided at position 5, and an antibody (hereinafter referred to as "immobilized antibody") that specifically reacts with the antigen to be measured in the sample at a different position is immobilized on the chromatographic medium. A position (detection part), 6 is a colored magnetic particle, 7 is a magnet (rubber magnet or the like), 8 is an enzyme chromogenic substrate (enzyme), and 9 is an immobilized enzyme (enzyme chromogenic substrate).
【００１８】図１において、測定対象抗原を含有する検
体を試料採取部３に施すと、毛管移動によって該抗原を
含む検体が下流側へと移動してゆき、位置４で標識抗体
と抗原・抗体反応を生じて標識された反応物を生ずる。
この標識された反応物はクロマトグラフ媒体上を更に下
流側へと毛管移動してゆき、検出部５で固定化抗体と更
に抗原・抗体反応を生じて検出部５に捕捉されて標識に
よる発現を行って、測定対象抗原の有無等の検出が行わ
れる。検体の残部はその後も下流側へと毛管移動を続
け、有色磁性粒子位置６に達し、該有色磁性粒子を随伴
しながら更に下流側に移動して磁石７位置に到達し、そ
こで磁石７によって捕捉されて有色磁性粒子に色に基づ
く呈色を発現して、試験の終了、すなわち検体がクロマ
トグラフ媒体の下流終端またはその付近に到達したこと
を明確に表示する。In FIG. 1, when a sample containing the antigen to be measured is applied to the sample collecting section 3, the sample containing the antigen moves to the downstream side by capillary movement, and at the position 4, the labeled antibody and the antigen / antibody The reaction occurs to give a labeled reactant.
The labeled reaction product migrates to the downstream side of the chromatographic medium by capillaries, and an antigen-antibody reaction occurs with the immobilized antibody in the detection unit 5 and is captured by the detection unit 5 to cause expression by the label. Then, the presence or absence of the antigen to be measured is detected. The rest of the sample continues capillary movement to the downstream side after that, reaches the position 6 of the colored magnetic particles, moves further downstream while accompanying the colored magnetic particles, and reaches the position of the magnet 7, where it is captured by the magnet 7. The colored magnetic particles are caused to develop a color based on the color to clearly indicate the end of the test, that is, the arrival of the sample at or near the downstream end of the chromatographic medium.
【００１９】図１において、磁石７は、検出試験装置と
一体に設けておいても、または着脱自在に設けておいて
も、或いは個別に準備しておき各々の試験内容等に応じ
て試験時に適当な位置に取り付けるようにしてもよい。
また、その場合に使用する有色磁性体の例としては、磁
性を有する着色したポリマー粒子；鉄、コバルト、ニッ
ケルまたはそれらの化合物等からなる着色した磁性物質
自体；体内にマグネタイトを持っている着色標識した細
菌（例えば走磁性細菌）等を挙げることができる。In FIG. 1, the magnet 7 may be provided integrally with the detection test apparatus, may be detachably provided, or may be separately prepared and may be prepared at the time of the test in accordance with each test content. It may be attached at an appropriate position.
In addition, examples of the colored magnetic substance used in that case include colored polymer particles having magnetism; a colored magnetic substance itself made of iron, cobalt, nickel or a compound thereof; a colored label having magnetite in the body. Examples of the bacteria include (eg, magnetotactic bacteria).
【００２０】また、図２においては、図１の場合と同様
に、試料採取部３に供給された測定対象抗原を含有する
検体が毛管移動によって下流側へと移動してゆき、位置
４で標識抗体と抗原・抗体反応を生じて標識された反応
物を生じ、この標識された反応物が検出部５で固定化抗
体と更に抗原・抗体反応を生じて検出部５に捕捉されて
標識により発現して、測定対象抗原の有無等の検出が行
われる。検体の残部はその後も下流側へと毛管移動を続
けて、８の位置の酵素発色性基質（酵素）に達し、その
後該酵素発色性基質（酵素）を随伴しながら更に下流側
に毛管移動して、固定化された酵素（酵素発色性基質）
９に到達し、そこで反応を生じて発色または色変化を起
こし、試験の終了を明確に表示する。Further, in FIG. 2, as in the case of FIG. 1, the sample containing the antigen to be measured supplied to the sample collecting unit 3 moves to the downstream side by capillary movement, and is labeled at the position 4. An antibody-antigen-antibody reaction is caused to generate a labeled reaction product, and this labeled reaction product further causes an antigen-antibody reaction with the immobilized antibody at the detection unit 5 to be captured by the detection unit 5 and expressed by the label. Then, the presence or absence of the antigen to be measured is detected. The rest of the sample continues capillary migration to the downstream side until it reaches the enzyme chromogenic substrate (enzyme) at the position 8 and then further migrates to the downstream side with the enzyme chromogenic substrate (enzyme). Immobilized enzyme (enzyme chromogenic substrate)
9 is reached, where a reaction takes place causing a color development or color change, clearly indicating the end of the test.
【００２１】図２のものでは、酵素発色性基質および酵
素の組合せの外に、酵素／酵素基質／発色性成分の組み
合わせを使用してもよい。その場合には、酵素、酵素基
質および発色性成分の１つまたは２つを所定の同じ位置
または別々の位置に施し、残りの２つまたは１つの成分
を毛管移動媒体の下終端またはその付近に固定して設け
るのがよい。その際の固定化は、この種の物質に対して
通常使用されている固定化方法を採用するのがよい。In FIG. 2, in addition to the enzyme chromogenic substrate and enzyme combination, the enzyme / enzyme substrate / chromogenic component combination may be used. In that case, one or two of the enzyme, the enzyme substrate and the chromogenic component are applied at the same predetermined position or different positions, and the remaining two or one components are placed at or near the lower end of the capillary migration medium. It is better to fix it. For immobilization at that time, it is preferable to adopt an immobilization method that is usually used for this type of substance.
【００２２】反応性発色剤である酵素発色性基質／酵素
の組合わせの例としては、アルカリホスファターゼ／５
−クロロー４−ブロモ−３−インドホスフェートからな
る組合せ、β−Ｄ−ガラクトシダーゼ／２−ニトロフェ
ニル・β−Ｄ−ガラクトシドからなる組合わせ等を挙げ
ることができる。また、反応性発色剤である酵素／酵素
基質／発色性成分の組合せの例としては、ペルオキシダ
ーゼ／ヒドロペルオキシド／２，２’−アジノビス（３
−エチルベンゾチアゾリン−６−スルホン酸）ジアンモ
ニウム塩（ＡＢＴＳ）からなる組合せ、グルコースオキ
シダーゼ／グルコース／ＡＢＴＳからなる組合せ等を挙
げることができる。An example of the combination of the enzyme chromogenic substrate / enzyme which is a reactive chromogenic agent is alkaline phosphatase / 5.
Examples include a combination of -chloro-4-bromo-3-indophosphate, a combination of β-D-galactosidase / 2-nitrophenyl / β-D-galactoside, and the like. Further, as an example of the combination of enzyme / enzyme substrate / color forming component which is a reactive color forming agent, peroxidase / hydroperoxide / 2,2′-azinobis (3
-Ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), a combination of glucose oxidase / glucose / ABTS, and the like.
【００２３】また、上記した酵素発色性基質／酵素の組
合せ、および酵素／酵素基質／発色性成分の組合せの代
わりに、通常の化学反応によって発色し得る複数の化合
物の組合せを使用してもよく、そのような例としては、
カリウム塩／コバルト亜硝酸ナトリウム、臭化物／硝酸
銀、硝酸銀／ジフェニルアミン、第１鉄塩／フェリシア
ン化カリウム、ヨウ素／デンプン等を挙げることができ
る。Further, instead of the above-mentioned enzyme chromogenic substrate / enzyme combination and enzyme / enzyme substrate / chromogenic component combination, a combination of a plurality of compounds capable of developing a color by an ordinary chemical reaction may be used. , As an example,
Examples thereof include potassium salt / sodium cobalt nitrite, bromide / silver nitrate, silver nitrate / diphenylamine, ferrous salt / potassium ferricyanide, iodine / starch and the like.
【００２４】更に、図１に示すものと図２に示すものを
組み合わせてもよく、その場合には、有色磁性粒子６の
位置と酵素発色性基質（酵素）８の位置は互いに離して
おいても、または互いに悪影響を及ぼさない時は同じ位
置にしてもよい。更に、磁石７の位置と酵素（酵素発色
性基質）９の位置は多少ずれしておいても、または同じ
位置にしておいてもよい。Further, the one shown in FIG. 1 and the one shown in FIG. 2 may be combined, in which case the positions of the colored magnetic particles 6 and the enzyme chromogenic substrate (enzyme) 8 are separated from each other. Or, they may be in the same position when they do not adversely affect each other. Further, the position of the magnet 7 and the position of the enzyme (enzyme color-developing substrate) 9 may be slightly shifted or may be the same position.
【００２５】図１および図２に示したものは、ほんの一
例であり、本発明はそれに限定されるものではない。そ
こで用いる毛管移動媒体等は、この種の検査試験におい
て従来から知られているいずれのものを使用してもよ
く、またそれらの形状もストリップ状だけでなくカラム
状等種々の形状とすることができる。The ones shown in FIGS. 1 and 2 are merely examples, and the present invention is not limited thereto. The capillary moving medium and the like used therefor may be any of those conventionally known in this type of inspection test, and the shape thereof may be various shapes such as column shape as well as strip shape. it can.
【００２６】そして、本発明においては、毛管移動媒体
の下流終端またはその付近における有色体用の捕捉材ま
たは反応型発色剤の該残りの成分の配置位置、毛管移動
媒体の材質や厚さ、毛管移動媒体の形状等を変えたり、
或いは毛管移動媒体は下流終端部に別の吸水体を設ける
等の方法を採用することによって、被検査流体が捕捉材
または反応型発色剤の該残りの成分の配置位置に到達す
るまでの時間、すなわち試験終了表示を発するまでの時
間を変化調節することができる。Further, in the present invention, the arrangement position of the remaining component of the capturing material for the colored body or the reactive color former at or near the downstream end of the capillary moving medium, the material and thickness of the capillary moving medium, the capillary Change the shape of the moving medium,
Alternatively, the capillary moving medium adopts a method such as providing another water absorbing body at the downstream end portion, whereby the fluid to be inspected reaches the arrangement position of the remaining component of the capturing material or the reactive color former, That is, it is possible to change and adjust the time until the test completion display is issued.
【００２７】そして、本発明は、毛管移動媒体を使用す
る検出試験法のいずれに対しても適用可能であり、例え
ば上記したイムノクロマトグラフ法のような免疫学的試
験法、毛管移動媒体を利用する化学分析等に有効に使用
できる。また、被検査流体は液体および気体のいずれの
形態であってもよい。以下に本発明を実施例により具体
的に説明するが、本発明はそれにより限定されない。The present invention can be applied to any of the detection test methods using a capillary transfer medium, and utilizes, for example, an immunological test method such as the immunochromatographic method described above, or a capillary transfer medium. It can be effectively used for chemical analysis. The fluid to be inspected may be in the form of either liquid or gas. Hereinafter, the present invention will be specifically described with reference to Examples, but the present invention is not limited thereto.
【００２８】[0028]
《実施例 １》試料採取部（親水性多孔性プラスチッ
ク；縦×横×厚さ＝５０ｍｍ×８ｍｍ×４ｍｍ）を上流
端（始端側）に有し、ポリエステルフイルムで裏打ちし
たナイロン・テトロン混紡織物（厚さ２５０μｍ）から
なる毛管移動媒体（幅８ｍｍ、長さ１００ｍｍ）を備え
たクロマトグラフを準備した。毛管移動媒体の始端から
１０ｍｍの位置に金コロイド粒子で標識された抗ヒト絨
毛性ゴナドトロピン（ＨＣＧ）ポリクロナール抗体の凍
結乾燥物１０μｇを施した。また、該標識抗体の位置か
ら１０ｍｍ離れた下流位置に抗ＨＣＧポリクロナール抗
体を固定化した。更に、該固定化抗体から１０ｍｍ下流
の位置に、青色磁性ポリスチレンラテックス粒子（固形
分１％；ポリサイエンスＩｎｃ.社製)３０μｌを施し、
また該青色磁性ポリスチレン粒子を施した位置から３０
ｍｍ下流（毛管移動媒体の始端から７０ｍｍの位置）
に、ゴム磁石（パイロット社製；マグシート；幅×長さ
×厚さ＝８ｍｍ×１ｍｍ×１ｍｍ）をポリエステル裏打
ちフイルム側に貼って、図１に示した検出試験具を作成
した。<< Example 1 >> A nylon / Tetron blended fabric having a sampling portion (hydrophilic porous plastic; length × width × thickness = 50 mm × 8 mm × 4 mm) at the upstream end (starting end side) and lined with a polyester film ( A chromatograph provided with a capillary moving medium (width: 8 mm, length: 100 mm) having a thickness of 250 μm) was prepared. 10 μg of a freeze-dried product of anti-human chorionic gonadotropin (HCG) polyclonal antibody labeled with gold colloid particles was applied at a position 10 mm from the beginning of the capillary migration medium. Further, an anti-HCG polyclonal antibody was immobilized at a downstream position 10 mm away from the position of the labeled antibody. Further, 30 μl of blue magnetic polystyrene latex particles (solid content 1%; manufactured by Polyscience Inc.) were applied at a position 10 mm downstream from the immobilized antibody,
In addition, 30 minutes from the position where the blue magnetic polystyrene particles are applied
mm downstream (position 70 mm from the start end of the capillary moving medium)
Then, a rubber magnet (manufactured by Pilot Co .; mug sheet; width × length × thickness = 8 mm × 1 mm × 1 mm) was attached to the polyester backing film side to prepare the detection test tool shown in FIG.
【００２９】上記で作成した検出試験具の試料採取部に
尿（検体）１ｍｌを浸み込ませて、イムノクロマトグラ
フ法による試験を行ったところ、抗ＨＣＧポリクロナー
ル抗体が固定化された箇所に検体中のＨＣＧが検出され
た。また、試験開始から３分後にゴム磁石位置に青色磁
性ポリスチレン粒子に基づく青色が観察されて、試験の
終了が呈色表示された。また、ゴム磁石の位置を毛管移
動媒体の始端から８０ｍｍに変えたところ、４分後に青
色呈色による試験終了表示がなされた。When 1 ml of urine (specimen) was soaked in the sampling part of the detection test device prepared above and a test by immunochromatography was conducted, it was found in the specimen that the anti-HCG polyclonal antibody was immobilized. HCG was detected. Further, after 3 minutes from the start of the test, a blue color based on the blue magnetic polystyrene particles was observed at the rubber magnet position, and the end of the test was displayed in color. Further, when the position of the rubber magnet was changed to 80 mm from the starting end of the capillary moving medium, after 4 minutes, the test completion display was made by blue coloring.
【００３０】《実施例 ２》実施例１で使用した検出試
験具の毛管移動媒体のゴム磁石を張り付けた下流終端部
分に濾紙(Bio-rad gel dryer;Bio-rad社製；縦５０ｍ
ｍ、横８ｍｍ、厚さ０.８５ｍｍ）を１枚または２枚重
ねて厚くし、且つゴム磁石の位置を毛管移動媒体の始端
から５５〜６５ｍｍの位置にした他は実施例１と同様に
して検出試験を行ったところ、試験終了を表示するまで
に要した時間は下記の表１に示すとおりであった。Example 2 A filter paper (Bio-rad gel dryer; manufactured by Bio-rad; length 50 m) is attached to the downstream end portion of the detection test tool used in Example 1 to which the rubber magnet of the capillary moving medium is attached.
m, width 8 mm, thickness 0.85 mm) by stacking one or two layers thicker, and the rubber magnet was positioned 55 to 65 mm from the starting end of the capillary moving medium, in the same manner as in Example 1. When a detection test was conducted, the time required to indicate the end of the test was as shown in Table 1 below.
【００３１】[0031]
【表１】 Bio-rad gel dryer枚数 ゴム磁石の位置 試験終了表示までの時間 (始端からの距離)) １ 枚 ６０ｍｍ ３分 ６５ｍｍ ４分 ２ 枚 ５５ｍｍ ３分 ６０ｍｍ ４分 [Table 1] Number of Bio-rad gel dryers Position of rubber magnet Time until display of test end (distance from start end ) 1 sheet 60mm 3 minutes 65mm 4 minutes 2 sheets 55mm 3 minutes 60mm 4 minutes
【００３２】上記表１の結果から、磁石の位置および／
または毛管移動媒体の状態（材質）等を変えることによ
って、試験終了を表示するのに要する時間を変化調節で
きることがわかる。From the results shown in Table 1 above, the magnet position and / or
It is also understood that the time required to display the end of the test can be changed and adjusted by changing the state (material) of the capillary moving medium.
【００３３】《実施例 ３》実施例１に使用した検出試
験具において、青色磁性ポリスチレン粒子を施す代わり
に、毛管移動媒体の始端から４５ｍｍの位置に２ｍＭ
ＭgＣｌ2−１０ｍＭ５−クロロー４−ブロモ−３−イン
ドホスフェートの１０μｌをスポイトで施し、また磁石
を設ける代わりに毛管移動媒体の始端から６０ｍｍの位
置にウシ腸由来のアルカリホスファターゼの３μｇを物
理吸着により固定化した検出試験具を使用した他は、実
施例１と同様にして検体の検出試験を行った。その結
果、抗ＨＣＧポリクロナール抗体が固定化された箇所に
検体中のＨＣＧが検出された。また、ウシ腸由来のアル
カリホスファターゼを固定化した位置に液が到達してか
ら３０〜６０秒後に青色の発色が明瞭に発現して試験の
終了を確認することができた。Example 3 In the detection test tool used in Example 1, instead of applying blue magnetic polystyrene particles, 2 mM was placed at a position 45 mm from the starting end of the capillary moving medium.
10 μl of MgCl 2 -10 mM 5-chloro-4-bromo-3-indophosphate was applied with a dropper, and instead of installing a magnet, 3 μg of alkaline phosphatase derived from calf intestine was fixed by physical adsorption at a position 60 mm from the beginning of the capillary transfer medium. A sample detection test was performed in the same manner as in Example 1 except that the modified detection test tool was used. As a result, HCG in the sample was detected at the location where the anti-HCG polyclonal antibody was immobilized. Further, 30 to 60 seconds after the liquid reached the position where the alkaline phosphatase derived from bovine intestine was immobilized, blue color was clearly developed, and the end of the test could be confirmed.
【００３２】《実施例 ４》実施例１に使用した検出試
験具において、青色磁性ポリスチレン粒子を施す代わり
に、毛管移動媒体の始端から４０ｍｍの位置にウレアハ
イドロジェンパーオキサイド（Sigma社製）１２.５μｇ
を施し、毛管移動媒体の始端から５０ｍｍの位置に西洋
ワサビ由来ペルオキシダーゼ２μｇを施し、また磁石を
設ける代わりに毛管移動媒体の始端から６０ｍｍの位置
にＡＢＴＳ４．５μｇを固定化した検出試験具を使用し
た他は実施例１と同様にして検体の検出試験を行った。
その結果、ＡＢＴＳを固定化した位置に液が到達してか
ら３０〜６０秒後に緑色の発色が明瞭に発現して試験の
終了を確認することができた。Example 4 Urea hydrogen peroxide (manufactured by Sigma) at the position 40 mm from the starting end of the capillary moving medium in the detection test tool used in Example 1 instead of applying the blue magnetic polystyrene particles. 5 μg
2 μg of horseradish-derived peroxidase was applied at a position 50 mm from the starting end of the capillary moving medium, and instead of providing a magnet, a detection test tool was used in which 4.5 μg of ABTS was immobilized at a position 60 mm from the starting end of the capillary moving medium. Others were the same as in Example 1, and the detection test of the sample was performed.
As a result, 30 to 60 seconds after the liquid reached the position where the ABTS was immobilized, the green color was clearly developed, and the end of the test could be confirmed.
【００３３】[0033]
【発明の効果】試験の終了時に鮮明な呈色状態または色
変化を発現させることが可能なため、試験の終了を明確
に表示することができる。毛管移動媒体および被検査流
体の色、種類、性状等に応じて有色体や反応型発色剤の
色や種類等を適宜選択することことができ、それによっ
て試験終了時の呈色状態を毛管移動媒体や被検査流体自
体の色と明確に区別できるようにすることが可能であ
る。 有色体や反応型発色剤を選択することによって、試験終
了表示部分を試験終了前の無色の状態から有色に発色さ
せることが可能であり、その場合には試験の終了が極め
て明確に表示できる。EFFECTS OF THE INVENTION Since it is possible to develop a clear coloration state or color change at the end of the test, it is possible to clearly indicate the end of the test. The color and type of the colored body or reactive color former can be appropriately selected according to the color, type and properties of the capillary moving medium and the fluid to be inspected. It is possible to make it possible to clearly distinguish it from the color of the medium or the fluid to be inspected. By selecting a colored body or a reactive color former, it is possible to color the test end display portion from a colorless state before the test to a color, and in that case, the end of the test can be displayed very clearly.
【図１】本発明の検出試験装置の一例を示す図である。FIG. 1 is a diagram showing an example of a detection test apparatus of the present invention.
【図２】本発明の検出試験装置の別の例を示す図であ
る。FIG. 2 is a diagram showing another example of the detection test apparatus of the present invention.
１ クロマトグラフ媒体 ２ 裏打ちフイルム ３ 試料採取部 ４ 標識抗体 ５ 固定化抗体 ６ 有色磁性粒子 ７ 磁石 ８ 酵素発色性基質（酵素） ９ 固定化された酵素（酵素発色性基質） 1 Chromatographic medium 2 Lining film 3 Sampling section 4 Labeled antibody 5 Immobilized antibody 6 Colored magnetic particles 7 Magnet 8 Enzyme chromogenic substrate (enzyme) 9 Immobilized enzyme (enzyme chromogenic substrate)
Claims (9)
特定物質を検出試験する方法において、毛管移動媒体の
所定位置に有色体を施すと共に、毛管移動媒体の下流終
端またはその付近に有色体用の捕捉材を位置させ、被検
査流体の毛管移動に伴って下流終端側に移動してきた有
色体を捕捉材で捕捉して試験の終了を呈色表示すること
を特徴とする方法。1. A method for detecting and testing a specific substance in a fluid to be inspected using a capillary moving medium, wherein a colored body is provided at a predetermined position of the capillary moving medium, and a colored substance is provided at or near a downstream end of the capillary moving medium. A method for locating a body-capturing material, capturing the colored body that has moved to the downstream end side with the capillary movement of the fluid to be inspected with the capturing material, and displaying the end of the test in color.
色体用の捕捉材が磁場発生部材である請求項１の方法。2. The method according to claim 1, wherein the colored body is colored magnetic particles, and the capturing material for the colored body is a magnetic field generating member.
特定物質を検出試験する方法において、毛管移動媒体の
所定位置に反応型発色剤の１つまたは２つ以上の成分を
施すと共に、毛管移動媒体の下流終端またはその付近に
反応型発色剤の残りの成分を位置させ、被検査流体の毛
管移動に伴って下流終端側に移動してきた反応型発色剤
の該１つまたは２つ以上の成分を該残りの成分と反応・
発色させて試験の終了を呈色表示することを特徴とする
方法。3. A method for detecting and testing a specific substance in a fluid to be tested using a capillary transfer medium, which comprises applying one or more components of a reactive color former to a predetermined position of the capillary transfer medium, The remaining components of the reactive color former are located at or near the downstream end of the capillary transfer medium, and the one or more reactive color formers that have moved to the downstream end side with the capillary movement of the fluid to be tested. React with the rest of the ingredients
A method characterized by displaying the color of the test to indicate the end of the test.
成分が酵素発色性基質または酵素であり、残りの成分が
それに対応した酵素または酵素発色性基質である請求項
３の方法。4. The method according to claim 3, wherein the one or more components of the reactive color former are enzyme color-forming substrates or enzymes, and the remaining components are corresponding enzymes or enzyme color-forming substrates.
る請求項３の方法。5. The method according to claim 3, wherein the reactive color former is a chemically reactive color former.
を伴う発色剤である請求項３の方法。6. The method according to claim 3, wherein the reactive color former is a color former involving a chemical reaction and an enzymatic reaction.
反応型発色成分の組み合わせからなる請求項６の方法。7. The method according to claim 6, wherein the reactive color former comprises a combination of an enzyme, an enzyme substrate and a reactive color developing component.
わせた方法。8. A method combining the methods of claim 1 and claim 3.
色体および／或いは反応型発色剤の１つまたは２つ以上
の成分、並びに毛管移動媒体の下流終端またはその付近
に配置した有色体用の捕捉材および／または反応型発色
剤の残りの成分を備えていることを特徴とする試験終了
表示体付きの検出試験装置。9. A capillary transfer medium, one or more components of a colored body and / or a reactive color developing agent applied at a predetermined position thereof, and a colored body arranged at or near the downstream end of the capillary transfer medium. A detection test device with a test completion indicator, characterized in that it comprises the remaining components of the scavenger and / or the reactive color former.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP24021791A JPH0552836A (en) | 1991-08-28 | 1991-08-28 | Displaying method for completion of test and detection tester fitted with test completion display |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP24021791A JPH0552836A (en) | 1991-08-28 | 1991-08-28 | Displaying method for completion of test and detection tester fitted with test completion display |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0552836A true JPH0552836A (en) | 1993-03-02 |
Family
ID=17056195
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP24021791A Pending JPH0552836A (en) | 1991-08-28 | 1991-08-28 | Displaying method for completion of test and detection tester fitted with test completion display |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0552836A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6194225B1 (en) | 1997-09-18 | 2001-02-27 | Matsushita Electric Industrial Co., Ltd. | Immunochromatography-assisted device |
| WO2002037108A1 (en) * | 2000-10-27 | 2002-05-10 | Morinaga Milk Industry Co., Ltd. | Reagent and method for detecting substance |
| EP1226434A1 (en) * | 1999-10-15 | 2002-07-31 | Wavesense, Llc | Systems and methods for performing magnetic chromatography assays |
| JP2012189453A (en) * | 2011-03-10 | 2012-10-04 | Toppan Printing Co Ltd | Detection method of substance to be detected using labeled magnetic particle, and detection system of substance to be detected |
-
1991
- 1991-08-28 JP JP24021791A patent/JPH0552836A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6194225B1 (en) | 1997-09-18 | 2001-02-27 | Matsushita Electric Industrial Co., Ltd. | Immunochromatography-assisted device |
| EP1226434A1 (en) * | 1999-10-15 | 2002-07-31 | Wavesense, Llc | Systems and methods for performing magnetic chromatography assays |
| EP1226434A4 (en) * | 1999-10-15 | 2005-04-06 | Wavesense Llc | Systems and methods for performing magnetic chromatography assays |
| NO331708B1 (en) * | 1999-10-15 | 2012-03-05 | Wavesense Llc | Systems and methods for performing magnetic chromatography analyzes |
| WO2002037108A1 (en) * | 2000-10-27 | 2002-05-10 | Morinaga Milk Industry Co., Ltd. | Reagent and method for detecting substance |
| JP2002202310A (en) * | 2000-10-27 | 2002-07-19 | Morinaga Milk Ind Co Ltd | Substance detecting reagent and substance detecting method |
| US7090984B2 (en) | 2000-10-27 | 2006-08-15 | Morinaga Milk Industry Co., Ltd. | Device and method for detecting substance |
| JP2012189453A (en) * | 2011-03-10 | 2012-10-04 | Toppan Printing Co Ltd | Detection method of substance to be detected using labeled magnetic particle, and detection system of substance to be detected |
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