JP5162439B2 - Immunochromatographic sensor - Google Patents
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- JP5162439B2 JP5162439B2 JP2008335838A JP2008335838A JP5162439B2 JP 5162439 B2 JP5162439 B2 JP 5162439B2 JP 2008335838 A JP2008335838 A JP 2008335838A JP 2008335838 A JP2008335838 A JP 2008335838A JP 5162439 B2 JP5162439 B2 JP 5162439B2
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- 239000000126 substance Substances 0.000 claims description 150
- 238000012800 visualization Methods 0.000 claims description 76
- 239000012085 test solution Substances 0.000 claims description 40
- 239000003153 chemical reaction reagent Substances 0.000 claims description 29
- 238000011161 development Methods 0.000 claims description 23
- HCAJQHYUCKICQH-VPENINKCSA-N 8-Oxo-7,8-dihydro-2'-deoxyguanosine Chemical compound C1=2NC(N)=NC(=O)C=2NC(=O)N1[C@H]1C[C@H](O)[C@@H](CO)O1 HCAJQHYUCKICQH-VPENINKCSA-N 0.000 claims description 20
- 239000000243 solution Substances 0.000 claims description 20
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 17
- 238000010521 absorption reaction Methods 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 12
- 239000000427 antigen Substances 0.000 claims description 9
- 108091007433 antigens Proteins 0.000 claims description 9
- 102000036639 antigens Human genes 0.000 claims description 9
- 239000012488 sample solution Substances 0.000 claims description 8
- 238000003892 spreading Methods 0.000 claims description 6
- 239000002245 particle Substances 0.000 claims description 4
- 238000004040 coloring Methods 0.000 claims description 2
- 239000004816 latex Substances 0.000 claims description 2
- 229920000126 latex Polymers 0.000 claims description 2
- 239000000049 pigment Substances 0.000 claims description 2
- 210000002700 urine Anatomy 0.000 description 21
- 238000006243 chemical reaction Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 230000036542 oxidative stress Effects 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 239000000463 material Substances 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 230000035882 stress Effects 0.000 description 3
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
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- 238000012790 confirmation Methods 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 230000000391 smoking effect Effects 0.000 description 2
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 description 1
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 description 1
- 239000006096 absorbing agent Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 229940029575 guanosine Drugs 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
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Description
本発明は、測定すべき物質と特異的に反応する抗体に金コロイド粒子等の可視化物質を結合させて、測定すべき物質の量を測定するイムノクロマトセンサに関する。 The present invention relates to an immunochromatographic sensor for measuring the amount of a substance to be measured by binding a visualization substance such as gold colloid particles to an antibody that specifically reacts with the substance to be measured.
「酸化ストレス状態」とは、体内で活性酸素と抗酸化物質とのバランスが崩れて活性酸素が優位になっている状態をいう。
喫煙、睡眠不足等の身体への過剰の負荷、過度のストレスが原因で抗酸化作用が低下すると、活性酸素が増加して酸化ストレスが高い状態になる。
酸化ストレスが高い状態は、上記したように、喫煙、睡眠不足等の身体への過剰の負荷、過度のストレスが原因で陥る症状であるため、これが長く続くと何らかの病気に進行していく可能性がある。また、体内の細胞の酸化が進むと身体が老化するという問題もある。
酸化ストレスが高い状態になると、遺伝子を構成する核酸塩基の一つであるグアノシンが酸化損傷され、細胞外に排出され、そして血液中、尿中に排出されることが知られている。
このため、血液や尿等の体液中の8-ヒドロキシデオキシグアノシンの量を測定することにより、病気になる前の未病の状態で病気になる可能性を発見することができる。
“Oxidative stress state” refers to a state in which active oxygen is dominant because the balance between active oxygen and antioxidants is lost in the body.
When the antioxidant action decreases due to excessive load on the body such as smoking and lack of sleep and excessive stress, active oxygen increases and oxidative stress becomes high.
As described above, a state with high oxidative stress is a symptom caused by excessive stress on the body such as smoking and lack of sleep, and excessive stress, so if this continues for a long time, it may progress to some kind of disease There is. Another problem is that the body ages as cells in the body oxidize.
It is known that when oxidative stress is high, guanosine, which is one of the nucleobases constituting the gene, is oxidatively damaged, excreted extracellularly, and excreted in blood and urine.
For this reason, by measuring the amount of 8-hydroxydeoxyguanosine in body fluids such as blood and urine, it is possible to discover the possibility of getting sick in an unaffected state before getting sick.
上記した8-ヒドロキシデオキシグアノシンのように１：１の抗体抗原反応しかすることができない低分子物質の量を手軽に測定する方法として、出願人は、測定すべき物質の抗体に、その抗体の存在を可視化できる可視化物質を結合させた可視化物質結合抗体を保持した可視化物質結合抗体層と、試液中の前記測定すべき物質と前記可視化物質結合抗体とが抗原抗体反応するのに充分な大きさの展開層とでテストストリップを構成し、前記展開層に、前記試液内の測定すべき物質と結合しなかった可視化物質結合抗体を結合させる抗原が固定された前記判定部を設けて成り、試液が、毛細管現象により、可視化物質結合抗体層を通って展開層に入り、展開層で試液中の前記測定すべき物質と前記可視化物質結合抗体とが抗体抗原反応した後、判定部を通過するように各層を配置したイムノクロマトセンサを提案している。
このイムノクロマトセンサを用いれば、測定すべき物質と結合しなかった可視化物質結合抗体が判定部の抗原に結合されて保持されるので、その可視化物質の量によって試液中の測定すべき物質の量を測定することができる。
即ち、測定すべき物質の量が多ければ多い程、可視化物質結合抗体層中の可視化物質結合抗体が測定すべき物質に結合されるため判定部で捕捉される可視化物質結合抗体の量は少なくなり、結果として判定部の色は薄くなり、逆に測定すべき物質の量が少なければ少ない程、判定部の色は濃くなる。
しかし、上記したイムノクロマトセンサは、試液が、毛細管現象により、可視化物質結合抗体層を通って展開層に入り、展開層で試液中の前記測定すべき物質と前記可視化物質結合抗体とが抗体抗原反応した後、判定部を通過するように構成されているため、例えば、何らかの詰まり等が原因で試液が可視化物質結合抗体層から展開層に入っていかないトラブルが発生する可能性がある。このようなトラブルが発生した場合、単に測定ができないだけなら他のセンサを用いてもう一回測定を行えば問題は解決するが、トラブルが発生していることに気付かないと、実際には試液が判定部に到達していないから判定部に色が付いていないにも拘わらず、判定部に色が付いていないから測定すべき物質の量が極めて多いと勘違いしてしまう可能性があるので問題である。
上記した問題は、出願人が提案した低分子物質用のイムノクロマトセンサに限らず、例えば、判定部で、可視化物質結合抗体と結合した測定すべき物質そのものを捕捉するように構成されているイムノクロマトセンサにも同様の問題が生じる。
出願人は、上記した問題点を見出し、本発明を発明するに至った。
本発明は、試液が、可視化物質結合抗体層を通って展開層に入ったことを容易に確認することができるイムノクロマトセンサを提供することを目的としている。
As a method for easily measuring the amount of a low molecular weight substance capable of performing only 1: 1 antibody antigen reaction, such as the above-mentioned 8-hydroxydeoxyguanosine, the applicant has applied the antibody of the substance to be measured to the antibody of the substance to be measured. A visualization substance-binding antibody layer holding a visualization substance-binding antibody bound with a visualization substance capable of visualizing the presence, and a size sufficient for an antigen-antibody reaction between the substance to be measured and the visualization substance-binding antibody in a test solution A test strip, and the development layer is provided with the determination unit on which an antigen that binds a visualization substance-bound antibody that did not bind to a substance to be measured in the test solution is fixed. However, due to capillarity, after entering the development layer through the visualization substance binding antibody layer, the substance to be measured in the test solution and the visualization substance binding antibody in the development layer after the antibody antigen reaction, It proposes the immunochromatographic sensor disposed each layer so as to pass through the tough.
If this immunochromatographic sensor is used, since the visualized substance-bound antibody that has not bound to the substance to be measured is bound and held by the antigen in the determination part, the amount of the substance to be measured in the test solution is determined by the amount of the visualized substance. Can be measured.
That is, the greater the amount of substance to be measured, the smaller the amount of visualization substance-bound antibody captured by the determination unit because the visualization substance-bound antibody in the visualization substance-binding antibody layer is bound to the substance to be measured. As a result, the color of the determination unit becomes lighter. Conversely, the smaller the amount of substance to be measured, the darker the color of the determination unit.
However, in the immunochromatographic sensor described above, the test solution enters the development layer through the visualization substance-binding antibody layer by capillary action, and the substance to be measured in the test solution and the visualization substance-binding antibody react with each other in the development layer. After that, since it is configured to pass through the determination unit, for example, there may be a problem that the test solution does not enter the developing layer from the visualization substance-bound antibody layer due to some clogging or the like. If such a trouble occurs, if the measurement cannot be performed simply, another measurement with another sensor will solve the problem, but if you do not notice that the trouble has occurred, However, because the determination part is not colored because it has not reached the determination part, the determination part is not colored. It is a problem.
The above-mentioned problem is not limited to the immunochromatographic sensor for low-molecular substances proposed by the applicant. For example, an immunochromatographic sensor configured to capture the substance to be measured itself bound to the visualized substance-binding antibody in the determination unit. A similar problem occurs.
The applicant has found the above problems and has invented the present invention.
An object of the present invention is to provide an immunochromatographic sensor capable of easily confirming that a test solution has entered a developing layer through a visualization substance-bound antibody layer.
上記した目的を達成するために本発明に係るイムノクロマトセンサは、測定すべき物質を含む試液を導入する試液吸収部と、測定すべき物質の抗体に、その存在を可視化できる物質を結合させた可視化物質結合抗体を含む可視化物質結合抗体層、及び試液中の前記測定すべき物質と、前記可視化物質結合抗体とが、抗原抗体反応するのに充分な大きさの展開層を備えたテストストリップと、前記テストストリップを収容するハウジングとを有し、前記展開層に、前記試液内の測定すべき物質と結合しなかった可視化物質結合抗体又は、前記試液内の測定すべき物質と結合した可視化物質結合抗体を結合させる抗原が固定された前記判定部が設けられ、試液が、毛細管現象により、可視化物質結合抗体層を通って展開層に入り、展開層で試液中の前記測定すべき物質と前記可視化物質結合抗体とが抗体抗原反応した後、判定部を通過するように各層が配置され、前記ハウジングに、少なくとも、前記判定部を外部から目視することができる窓部と、前記可視化物質結合抗体層の少なくとも一部を外部から目視することができる窓部とを設けたことを特徴とする。
好ましくは、前記前記判定部を外部から目視することができる窓部と、前記可視化物質結合抗体層の少なくとも一部を外部から目視することができる窓部とは一つの窓部で形成され得る。
また、前記試液吸収部は、別体の試液採取パッドで採取した試液を含浸させることができるように構成してもよく、滴下した試液を含浸させることができるように構成されていてもよく、試液を注入することができるように構成されていてもよい。
前記測定すべき物質は、例えば、8-ヒドロキシデオキシグアノシンであり、測定すべき物質の抗体が8OHdG1B1(独立行政法人 産業技術総合研究所 特許生物寄託センター 受託番号FERM P-20122)（本願の出願人が特開2006-056859にて既に提案している抗体）であり得る。
また、前記可視化物質は、例えば、金コロイド等の有色物質であり得る。
さらにまた、前記可視化物質は、ラテックス粒子、着色色素又は蛍光物質の何れか一つであり得る。
In order to achieve the above-described object, the immunochromatographic sensor according to the present invention includes a reagent solution absorption part for introducing a reagent solution containing a substance to be measured, and a visualization in which a substance capable of visualizing the presence is bound to an antibody of the substance to be measured. A visualization substance-binding antibody layer containing a substance-binding antibody, and a test strip provided with a development layer that is large enough for antigen-antibody reaction between the substance to be measured in the test solution and the visualization substance-binding antibody; A housing containing the test strip, and a visualized substance-binding antibody that has not bound to the substance to be measured in the test solution or a visualized substance binding to the developing layer bound to the substance to be measured in the test solution. The determination unit to which the antigen to which the antibody is bound is fixed is provided, and the test solution enters the development layer through the visualization substance binding antibody layer by capillary action, and in the test solution in the development layer. After the substance to be measured and the visualization substance-bound antibody have reacted with each other, each layer is disposed so as to pass through the determination unit, and at least the determination unit can be visually observed from the outside in the housing. And a window portion through which at least a part of the visualized substance-binding antibody layer can be visually observed from the outside.
Preferably, the window part from which the determination part can be visually observed from the outside and the window part from which at least a part of the visualized substance-binding antibody layer can be visually observed from the outside can be formed by one window part.
Further, the reagent solution absorption part may be configured to be able to impregnate a sample solution collected by a separate sample solution collecting pad, or may be configured to be able to impregnate a dropped test solution, You may be comprised so that a test solution can be inject | poured.
The substance to be measured is, for example, 8-hydroxydeoxyguanosine, and the antibody of the substance to be measured is 8OHdG1B1 (National Institute of Advanced Industrial Science and Technology, Patent Biodeposition Center Accession No.FERM P-20122) (the applicant of the present application). Can be the antibody already proposed in JP-A-2006-056859.
The visualization material may be a colored material such as gold colloid, for example.
Furthermore, the visualization material may be any one of latex particles, coloring pigments, or fluorescent materials.
本発明に係るイムノクロマトセンサは、測定すべき物質を含む試液を導入する試液吸収部と、測定すべき物質の抗体に、その存在を可視化できる物質を結合させた可視化物質結合抗体を含む可視化物質結合抗体層、及び試液中の前記測定すべき物質と、前記可視化物質結合抗体とが、抗原抗体反応するのに充分な大きさの展開層を備えたテストストリップと、前記テストストリップを収容するハウジングとを有し、前記展開層に、前記試液内の測定すべき物質と結合しなかった可視化物質結合抗体又は、前記試液内の測定すべき物質と結合した可視化物質結合抗体を結合させる抗原が固定された前記判定部が設けられ、試液が、毛細管現象により、可視化物質結合抗体層を通って展開層に入り、展開層で試液中の前記測定すべき物質と前記可視化物質結合抗体とが抗体抗原反応した後、判定部を通過するように各層が配置され、前記ハウジングに、少なくとも、前記判定部を外部から目視することができる窓部と、前記可視化物質結合抗体層の少なくとも一部を外部から目視することができる窓部とを設けているので、試液が通過する時に可視化物質結合抗体層に保持された可視化物質結合抗体が試液中に溶出することによる可視化物質結合抗体層の色の変化によって、試液が、可視化物質結合抗体層を通って展開層に入ったか否かを目視確認することができるようになる。
また、このように可視化物質結合抗体層の色の変化を利用して試液の流れ具合を確認することができるように構成することにより、試液の流れ具合を確認するための特別な構成を必要としないので、構造が簡単であり、安価に製造できるという効果も奏する。
結果として、試液の詰まり等のトラブルに気付かずに、判定結果を見誤るという問題がなくなる。
また、前記前記判定部を外部から目視することができる窓部と、前記可視化物質結合抗体層の少なくとも一部を外部から目視することができる窓部とは一つの窓部で形成することにより、ハウジングの加工が容易であるという効果を奏する。
前記試液吸収部を、別体の試液採取パッドで採取した試液を含浸させることができるように構成することにより、試液吸収部に直接試液をかける必要がなくなるので、試液の採取が容易である。
また、前記測定すべき物質を、8-ヒドロキシデオキシグアノシンとし、測定すべき物質の抗体を8OHdG1B1(独立行政法人 産業技術総合研究所 特許生物寄託センター 受託番号FERM P-20122)（本願の出願人が特開2006-056859にて既に提案している抗体）とすることによって、「酸化ストレス状態」を手軽に測定することが可能になる。
The immunochromatographic sensor according to the present invention includes a reagent absorption part that introduces a reagent solution containing a substance to be measured, and a visualization substance binding that includes a visualization substance-binding antibody in which a substance that can be visualized is bound to an antibody of the substance to be measured. An antibody layer, a test strip provided with a development layer large enough to cause an antigen-antibody reaction between the substance to be measured in the test solution and the visualization substance-bound antibody, and a housing for housing the test strip And an antigen that binds the visualization substance-bound antibody that has not bound to the substance to be measured in the test solution or the visualization substance-bound antibody that has bound to the substance to be measured in the test solution is fixed to the spreading layer. The determination unit is provided, and the test solution enters the development layer through the visualization substance-bound antibody layer by capillary action, and the substance to be measured in the test solution and the possible substance are detected in the development layer. After the antigen-antibody reaction with the visualization substance-bound antibody, each layer is arranged so as to pass through the judgment part, and at least the judgment part can be visually observed from the outside in the housing, and the visualization substance-binding antibody Since a window part is provided so that at least a part of the layer can be visually observed from the outside, a visualization substance is obtained by elution of the visualization substance-bound antibody retained in the visualization substance-binding antibody layer when the reagent passes through The change in the color of the bound antibody layer makes it possible to visually check whether the test solution has entered the spreading layer through the visualization substance-bound antibody layer.
In addition, a special configuration for confirming the flow of the reagent solution is required by using the change in the color of the visualized substance binding antibody layer so that the flow of the reagent solution can be confirmed. Therefore, the structure is simple, and there is an effect that it can be manufactured at low cost.
As a result, there is no problem of mistaking the determination result without noticing trouble such as clogging of the test solution.
In addition, by forming the window that can be seen from the outside and the window that can be seen from the outside at least a part of the visualized substance-bound antibody layer by forming a single window, There is an effect that the processing of the housing is easy.
By configuring the reagent solution absorption part to be impregnated with a test solution collected by a separate sample solution collection pad, it is not necessary to apply the test solution directly to the reagent solution absorption part, so that the sample solution can be collected easily.
The substance to be measured is 8-hydroxydeoxyguanosine, and the antibody of the substance to be measured is 8OHdG1B1 (National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Accession No.FERM P-20122) (the applicant of the present application By using the antibody already proposed in Japanese Patent Laid-Open No. 2006-056859, the “oxidative stress state” can be easily measured.
以下、添付図面に示した一実施例を参照して、本発明に係るイムノクロマトセンサの実施の形態を説明していく。 Embodiments of an immunochromatographic sensor according to the present invention will be described below with reference to one embodiment shown in the accompanying drawings.
図１は本発明に係るイムノクロマトセンサの一実施例の分解展開図、図２は図１に示したイムノクロマトセンサの上面カバー以外の部分を組立てた部分分解展開図、図３は図１に示したイムノクロマトセンサの上面図を各々示している。
図中、符号１は基板を示しており、この基板１の上に、テストストリップ２及び試液吸収部３が設けられている。
テストストリップ２は、試液吸収部３に部分的に重ねて配置された試液展開層４と、前記試液展開層４に部分的に重ねて配置された可視化物質結合抗体保持層５と、標識保持層５に部分的に重ねて配置された展開層６とから成る。
可視化物質結合抗体保持層５には、測定すべき物質と特異的に反応する抗体に、その抗体の存在を可視化できる可視化物質を結合させた可視化物質結合抗体が保持されている。具体的には、例えば、測定すべき物質が8-ヒドロキシデオキシグアノシンの場合、その抗体は、8OHdG1B1(独立行政法人 産業技術総合研究所 特許生物寄託センター 受託番号FERM P-20122)（本願の出願人が特開2006-056859にて既に提案している抗体））であり、可視化物質としては金コロイドが挙げられる。この可視化物質結合抗体保持層５に保持された可視化物質結合抗体は、試液展開層４を介して試液が導入されると試液中に溶出する。このため、可視化物質結合抗体保持層５は、使用前（即ち、試液が通過する前）は、可視化物質（例えば、金コロイド）によって色付いているが、使用後、即ち、試液が正常に通過した後は、可視化物質が試液中に溶出してしまうため色が無くなる。具体的には、可視化物質結合抗体保持層５が白いメンブレンから成り、可視化物質が金コロイドである場合には、可視化物質結合抗体保持層５は、使用前は赤色であるが、使用後は白色になる。
展開層６は、前記可視化物質結合抗体保持層５に保持された可視化物質結合抗体と結合して、同抗体を捕捉する固定化用物質が固定された判定部７を有する。この判定部７は、展開層６において測定すべき物質と可視化物質結合抗体とが充分に抗体抗原反応することができるように、可視化物質結合抗体保持層５から充分に離れた位置に設けられている。
1 is an exploded view of an embodiment of an immunochromatographic sensor according to the present invention, FIG. 2 is a partially exploded view in which parts other than the top cover of the immunochromatographic sensor shown in FIG. 1 are assembled, and FIG. 3 is shown in FIG. The top view of an immunochromatographic sensor is shown, respectively.
In the figure, reference numeral 1 denotes a substrate, on which a test strip 2 and a reagent solution absorbing portion 3 are provided.
The test strip 2 includes a reagent solution developing layer 4 that is partially overlapped with the reagent solution absorbing portion 3, a visualization substance-binding antibody holding layer 5 that is partially overlapped with the reagent solution developing layer 4, and a label holding layer. 5 and a development layer 6 that is partially overlapped with 5.
The visualization substance-bound antibody holding layer 5 holds a visualization substance-binding antibody in which a visualization substance capable of visualizing the presence of the antibody is bound to an antibody that specifically reacts with the substance to be measured. Specifically, for example, when the substance to be measured is 8-hydroxydeoxyguanosine, the antibody is 8OHdG1B1 (National Institute of Advanced Industrial Science and Technology, Patent Biodeposition Center Accession No.FERM P-20122) (the applicant of the present application). Is an antibody already proposed in Japanese Patent Application Laid-Open No. 2006-056859)), and examples of the visualization substance include colloidal gold. The visualized substance-bound antibody retained in the visualized substance-bound antibody retaining layer 5 is eluted in the reagent when the reagent is introduced through the reagent developing layer 4. Therefore, the visualization substance-bound antibody holding layer 5 is colored by the visualization substance (for example, gold colloid) before use (that is, before the sample solution passes), but after use, that is, the sample solution has passed normally. After that, since the visualization substance is eluted in the test solution, the color disappears. Specifically, when the visualization substance binding antibody holding layer 5 is made of a white membrane and the visualization substance is a gold colloid, the visualization substance binding antibody holding layer 5 is red before use but is white after use. become.
The development layer 6 has a determination unit 7 to which an immobilization substance that binds to the visualization substance-binding antibody held in the visualization substance-binding antibody holding layer 5 and captures the antibody is fixed. The determination unit 7 is provided at a position sufficiently separated from the visualization substance-bound antibody holding layer 5 so that the substance to be measured in the development layer 6 and the visualization substance-bound antibody can sufficiently react with each other. Yes.
上記したように構成されたテストストリップ２及び試液吸収部３は土台となるベース基板１上に配置され、さらにベース基板１にカバー１０が取り付けられる。
この実施例では、カバー１０は、ベース基板１における試液吸収部３以外の部分を全て覆うように寸法決めされており、かつ、テストストリップ２に相当する部分には確認用窓１１が形成されている。
この確認用窓１１は、可視化物質結合抗体保持層５の一部と、展開層６における判定部７が露出するように形成されている。
また、カバー１０における判定部７に相当する部分には、判定部７を挟むように色調表１２が設けられている。この実施例では、図３における判定部７の左側に測定すべき物質の量が少ない時の色（即ち、濃い色）が、右側に測定すべき物質の量が多い時の色（即ち、薄い色）が印刷されている。
The test strip 2 and the test solution absorber 3 configured as described above are disposed on the base substrate 1 serving as a base, and a cover 10 is attached to the base substrate 1.
In this embodiment, the cover 10 is dimensioned so as to cover all portions of the base substrate 1 other than the reagent solution absorbing portion 3, and a confirmation window 11 is formed in a portion corresponding to the test strip 2. Yes.
This confirmation window 11 is formed so that a part of the visualization substance binding antibody holding layer 5 and the determination part 7 in the development layer 6 are exposed.
A color tone table 12 is provided at a portion corresponding to the determination unit 7 in the cover 10 so as to sandwich the determination unit 7. In this embodiment, the color when the amount of the substance to be measured is small on the left side of the determination unit 7 in FIG. 3 (that is, the dark color) is the color when the amount of the substance to be measured is large on the right side (that is, the color is light). Color) is printed.
次に、上記したように構成されたイムノクロマトセンサの作用を説明していく。
本発明に係るイムノクロマトセンサの作用を示す図であり、（ａ）は使用前の状態を、（ｂ）は使用後の状態を各々示している。
この説明では、測定すべき物質は、尿中に含まれている8-ヒドロキシデオキシグアノシンの量であり、従って、抗体は、8-ヒドロキシデオキシグアノシンに特異的に反応する抗体（8OHdG1B1(独立行政法人 産業技術総合研究所 特許生物寄託センター 受託番号FERM P-20122)（本願の出願人が特開2006-056859にて既に提案している抗体）であり、可視化物質は金コロイドである。また、判定層に固定されている固定化物質は8-ヒドロキシデオキシグアノシン(8-OHdG)である。
使用者が、試液吸収部３に尿を直接かけるか、別体のスポイト（図示せず）等で試液吸収部３に尿を滴下するか、又は、予め尿をかけておいた別体の採尿パット８を試液吸収部３の上に置くことによって、尿は試液吸収部３に含浸する。試液吸収部３に含浸した尿は、毛細管現象により試液展開層４に入り、試液展開層４の幅全体に広がりながら可視化物質結合抗体保持層５に進む。そして、尿が可視化物質結合抗体保持層５を通過する時に、尿中に、同層５に保持された金コロイドが結合した抗体（8OHdG1B1(独立行政法人 産業技術総合研究所 特許生物寄託センター 受託番号FERM P-20122)（本願の出願人が特開2006-056859にて既に提案している抗体））が溶出する。金コロイド結合抗体が溶出した尿は、そのまま展開層６へ進み、同層６で8-ヒドロキシデオキシグアノシンと金コロイド結合抗体とが抗体抗原反応して、尿中にある8-ヒドロキシデオキシグアノシンに金コロイド結合抗体が結合する。展開層６に入った尿は、さらに、毛細管現象により展開層６を判定部７に向かって進む。
尿が判定部７を通過する時に、既に8-ヒドロキシデオキシグアノシンと結合した金コロイド結合抗体は判定部７を通過するが、8-ヒドロキシデオキシグアノシンと結合していない金コロイド結合抗体は判定部７に固定された固定化物質と結合して判定部７に留まる。このため、判定部７は固定化物質と結合した金コロイド結合抗体の金コロイドの色の作用で赤く見える。
尿中に含まれている8-ヒドロキシデオキシグアノシンの量が多ければ多いほど、8-ヒドロキシデオキシグアノシンと結合しない金コロイド結合抗体の量は少なくなるので判定部７の色は薄くなり、逆に、尿中に含まれている8-ヒドロキシデオキシグアノシンの量が少なければ少ないほど、8-ヒドロキシデオキシグアノシンと結合しない金コロイド結合抗体の量が多くなるので判定部７の色は濃くなる。
使用者は、判定部７の両隣にある色調表１２の色と判定部７の色とを比較して、尿中に含まれている8-ヒドロキシデオキシグアノシンの量が多いか否かを判断する。
ところで、上記したように、このイムノクロマトセンサはカバー１０の窓部１１が、可視化物質結合抗体保持層５の一部が露出するように形成されている。このため、使用者は、窓部１１から見える可視化物質結合抗体保持層５の色の変化によって、尿が展開層６に入っているか否かを確認することができる。具体的には、可視化物質結合抗体保持層５は、使用前は、そこに保持されている可視化物質結合抗体の可視化物質、ここでは金コロイドの作用で色付いている。そして、可視化物質結合抗体保持層５は試液、ここでは尿が通過すると、可視化物質結合抗体が試液中に溶出するため同層５の色はなくなる。従って、試液吸収部３から導入した尿が、何らかの原因で展開層６に進まなければ可視化物質結合抗体保持層５の色は変わらないが、試液吸収部３から導入した尿が、正確に展開層６に進めば可視化物質結合抗体保持層５の色が変わる。このため、使用者は、窓部１１から見える可視化物質結合抗体保持層５の色によって試液が正確に展開層６に進んでいるか否かを確認することができるため、判定結果を間違えて認識することがない。
Next, the operation of the immunochromatographic sensor configured as described above will be described.
It is a figure which shows the effect | action of the immunochromatographic sensor which concerns on this invention, (a) has shown the state before use, (b) has each shown the state after use.
In this description, the substance to be measured is the amount of 8-hydroxydeoxyguanosine contained in the urine. Therefore, the antibody is an antibody (8OHdG1B1 (independent administrative agency) that specifically reacts with 8-hydroxydeoxyguanosine. (National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center, Accession No. FERM P-20122) (An antibody already proposed by the applicant of the present application in Japanese Patent Laid-Open No. 2006-056859), and the visualization substance is a colloidal gold. The immobilizing substance immobilized on the layer is 8-hydroxydeoxyguanosine (8-OHdG).
The user puts urine directly on the test solution absorption part 3, or drops urine on the test solution absorption part 3 with a separate dropper (not shown), or separate urine collection previously applied with urine By placing the pad 8 on the reagent solution absorbing part 3, urine is impregnated in the reagent solution absorbing part 3. The urine impregnated in the test solution absorbing part 3 enters the test solution developing layer 4 by capillary action and proceeds to the visualization substance-bound antibody holding layer 5 while spreading over the entire width of the test solution developing layer 4. When the urine passes through the visualization substance-binding antibody holding layer 5, the antibody (8OHdG1B1 (Independent Administrative Institution National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Accession No.) FERM P-20122) (an antibody already proposed by the applicant of the present application in Japanese Patent Laid-Open No. 2006-056859). The urine from which the gold colloid-binding antibody has been eluted proceeds to the developing layer 6 as it is, and 8-hydroxydeoxyguanosine and the gold colloid-binding antibody react with the antibody antigen in the same layer 6 to convert 8-hydroxydeoxyguanosine in the urine into gold. Colloid binding antibody binds. The urine that has entered the development layer 6 further proceeds through the development layer 6 toward the determination unit 7 by capillary action.
When the urine passes through the determination unit 7, the gold colloid-binding antibody already bound to 8-hydroxydeoxyguanosine passes the determination unit 7, but the gold colloid-binding antibody not bonded to 8-hydroxydeoxyguanosine determines the determination unit 7. It remains in the determination unit 7 by combining with the immobilized substance fixed on the surface. For this reason, the determination unit 7 looks red due to the effect of the colloidal gold color of the colloidal gold antibody bound to the immobilized substance.
The greater the amount of 8-hydroxydeoxyguanosine contained in the urine, the smaller the amount of colloidal gold-bound antibody that does not bind to 8-hydroxydeoxyguanosine, so the color of the determination unit 7 becomes lighter. The smaller the amount of 8-hydroxydeoxyguanosine contained in the urine, the greater the amount of colloidal gold-bound antibody that does not bind to 8-hydroxydeoxyguanosine, so the color of the determination unit 7 becomes darker.
The user compares the color of the color tone table 12 on both sides of the determination unit 7 with the color of the determination unit 7 to determine whether the amount of 8-hydroxydeoxyguanosine contained in the urine is large. .
By the way, as described above, in this immunochromatographic sensor, the window portion 11 of the cover 10 is formed so that a part of the visualization substance binding antibody holding layer 5 is exposed. For this reason, the user can confirm whether or not urine is in the spreading layer 6 by a change in the color of the visualization substance binding antibody holding layer 5 visible from the window portion 11. Specifically, the visualized substance-bound antibody holding layer 5 is colored by the action of the visualized substance of the visualized substance-bound antibody held therein, here, colloidal gold, before use. The visualization substance-binding antibody holding layer 5 disappears when the test solution, here urine passes, because the visualization substance-binding antibody is eluted in the test solution. Therefore, the color of the visualization substance-bound antibody holding layer 5 does not change unless the urine introduced from the test solution absorption part 3 proceeds to the development layer 6 for some reason. If it advances to 6, the color of the visualization substance binding antibody holding layer 5 will change. For this reason, since the user can confirm whether or not the test solution has advanced to the developing layer 6 accurately based on the color of the visualization substance-binding antibody holding layer 5 that can be seen from the window portion 11, the determination result is mistakenly recognized. There is nothing.
上記した実施例では、判定部７において、測定すべき物質（本実施例では、8-ヒドロキシデオキシグアノシン）と結合しなかった可視化物質結合抗体を捕捉して、測定すべき物質の量を判定しているが、この構成は実施例に限定されることなく、測定すべき物質の種類に応じて、例えば、判定部において、測定すべき物質と結合した可視化物質結合抗体そのものを捕捉して、測定すべき物質の量を判定するように構成してもよい。 In the embodiment described above, the determination unit 7 captures the visualization substance-bound antibody that has not bound to the substance to be measured (in this example, 8-hydroxydeoxyguanosine), and determines the amount of the substance to be measured. However, this configuration is not limited to the embodiment, and depending on the type of the substance to be measured, for example, in the determination unit, the visualization substance-bound antibody itself bound to the substance to be measured is captured and measured. You may comprise so that the quantity of the substance which should be determined may be determined.
上記した実施例では、一つの窓部１１で、判定部７と可視化物質結合抗体保持層５との両方を見ることができるように構成しているため、ハウジングを構成するカバー１０の製造が容易であるという効果を奏するが、窓部１１の構成は本実施例に限定されることなく、例えば、図５に示すように、判定部７用の窓部１１ａと、可視化物質結合抗体保持層５用の窓部１１ｂとを別々に設けてもよい。 In the above-described embodiment, since the single window portion 11 is configured so that both the determination portion 7 and the visualization substance-bound antibody holding layer 5 can be seen, the manufacture of the cover 10 constituting the housing is easy. However, the configuration of the window 11 is not limited to the present embodiment. For example, as shown in FIG. 5, the window 11 a for the determination unit 7 and the visualization substance-bound antibody holding layer 5 are provided. The window portion 11b may be provided separately.
１ ベース基板
２ テストストリップ
３ 試液吸収部
４ 試液展開層
５ 可視化物質結合抗体保持層
６ 展開層
７ 判定部
８ 採尿パッド
１０ カバー
１１ 窓部
１１ａ 窓部
１１ｂ 窓部
１２ 色調表
DESCRIPTION OF SYMBOLS 1 Base substrate 2 Test strip 3 Reagent absorption part 4 Reagent development layer 5 Visualization substance binding antibody holding layer 6 Development layer 7 Judgment part 8 Urine collection pad 10 Cover 11 Window part 11a Window part 11b Window part 12 Color tone table
Claims (8)
測定すべき物質の抗体に、その存在を可視化できる物質を結合させた可視化物質結合抗体を含む可視化物質結合抗体層、及び試液中の前記測定すべき物質と、前記可視化物質結合抗体とが、抗原抗体反応するのに充分な大きさの展開層を備えたテストストリップと、
前記テストストリップを収容するハウジングと
を有し、
前記展開層に、
前記試液内の測定すべき物質と結合しなかった可視化物質結合抗体又は、
前記試液内の測定すべき物質と結合した可視化物質結合抗体
を結合させる抗原が固定された前記判定部が設けられ、
試液が、毛細管現象により、可視化物質結合抗体層を通って展開層に入り、展開層で試液中の前記測定すべき物質と前記可視化物質結合抗体とが抗体抗原反応した後、判定部を通過するように各層が配置され、
前記ハウジングに、少なくとも、前記判定部を外部から目視することができる窓部と、前記可視化物質結合抗体層の少なくとも一部を外部から目視することができる窓部とを設けた
ことを特徴とするイムノクロマトセンサ。 A reagent absorption part for introducing a reagent containing a substance to be measured;
A visualization substance-binding antibody layer including a visualization substance-binding antibody in which a substance capable of visualizing the presence of the substance is bound to an antibody of the substance to be measured, and the substance to be measured in the test solution and the visualization substance-binding antibody are antigens A test strip with a spreading layer large enough to react with the antibody;
A housing for housing the test strip;
In the spreading layer,
A visualized substance-bound antibody that did not bind to the substance to be measured in the test solution, or
The determination unit to which an antigen that binds the visualization substance-bound antibody bound to the substance to be measured in the test solution is fixed;
The test solution enters the development layer through the visualization substance-binding antibody layer by capillary action, and passes through the determination unit after the substance to be measured in the test solution and the visualization substance-binding antibody react with each other in the development layer. Each layer is placed as
The housing is provided with at least a window part through which the determination part can be visually observed from the outside and a window part through which at least a part of the visualization substance-binding antibody layer can be visually observed from the outside. Immunochromatographic sensor.
ことを特徴とする請求項１に記載のイムノクロマトセンサ。 A window part that allows the determination part to be visually recognized from the outside and a window part that enables at least a part of the visualization substance-bound antibody layer to be visually observed from the outside are formed by one window part. The immunochromatographic sensor according to claim 1.
ことを特徴とする請求項１又は２に記載のイムノクロマトセンサ。 The immunochromatographic sensor according to claim 1 or 2, wherein the reagent solution absorption unit is configured to be able to impregnate a sample solution collected by a separate sample solution collecting pad.
ことを特徴とする請求項１又は２に記載のイムノクロマトセンサ。 The immunochromatographic sensor according to claim 1 or 2, wherein the reagent solution absorbing section is configured to be able to impregnate the dropped reagent solution.
ことを特徴とする請求項１又は２に記載のイムノクロマトセンサ。 The immunochromatographic sensor according to claim 1, wherein the reagent solution absorption unit is configured to be able to inject a reagent solution.
ことを特徴とする請求項１〜５の何れか一項に記載のイムノクロマトセンサ。 The substance to be measured is 8-hydroxydeoxyguanosine, and the antibody of the substance to be measured is 8OHdG1B1 (National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary Accession No.FERM P-20122) The immunochromatographic sensor according to any one of claims 1 to 5.
ことを特徴とする請求項１〜６の何れか一項に記載のイムノクロマトセンサ。 The immunochromatographic sensor according to any one of claims 1 to 6, wherein the visualization substance is one of latex particles, a coloring pigment, and a fluorescent substance.
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