JPH11326326A - Immunity inspection body - Google Patents
Immunity inspection bodyInfo
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- JPH11326326A JPH11326326A JP13022298A JP13022298A JPH11326326A JP H11326326 A JPH11326326 A JP H11326326A JP 13022298 A JP13022298 A JP 13022298A JP 13022298 A JP13022298 A JP 13022298A JP H11326326 A JPH11326326 A JP H11326326A
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- unit
- immunoassay
- antibody
- immunoassays
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Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、抗原抗体反応によ
り被検体、特にスギ、ダニ等のアレルゲンを検出する免
疫検査体に関する。特に被検体の存在量範囲を評価する
ことのできる機能を組み込んだ免疫検査体に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an immunological test for detecting an allergen such as a test substance, particularly cedar or tick, by an antigen-antibody reaction. In particular, the present invention relates to an immunological test body incorporating a function capable of evaluating the range of abundance of a subject.
【０００２】[0002]
【従来の技術】対象抗原を検出する方法として、一般に
イムノクロマト法が用いられている。イムノクロマト法
では、対象抗原と結合する２種類の抗体、即ち標識抗体
と固定抗体とが組み込まれたクロマト基材の一端を、対
象抗原を溶解した溶液中に浸漬することによりクロマト
展開が行われる。ここで用いられる標識抗体とは、通常
金コロイド（赤）または着色ラテックス（各種の色が用
いられるが青が一般的）を付加したものであり、展開液
に溶解可能な形でクロマト基材に保持されており、固定
抗体とは標識抗体とは異なるエピトープで対象抗原と結
合する性質を持つもので、標識抗体の保持されている位
置よりも後方のクロマト基材上に固定されている。この
場合、通常は一つの対象抗原に対してただ１組の抗体対
が用いられる。2. Description of the Related Art As a method for detecting a target antigen, an immunochromatography method is generally used. In the immunochromatography method, one end of a chromatographic substrate in which two kinds of antibodies that bind to a target antigen, that is, a labeled antibody and a fixed antibody are incorporated, is immersed in a solution in which the target antigen is dissolved to perform chromatographic development. The labeled antibody used here is usually a substance to which gold colloid (red) or colored latex (various colors are used, but blue is generally used) is added to a chromatographic substrate in a form that can be dissolved in a developing solution. It is retained, and the immobilized antibody is an epitope different from that of the labeled antibody and has a property of binding to the target antigen, and is immobilized on the chromatographic base material behind the position where the labeled antibody is retained. In this case, usually only one set of antibody pairs is used for one antigen of interest.
【０００３】この従来のイムノクロマト法では対象抗
原、即ち被検出物の有無は検出部の呈色の有無によって
判定され、またその量は、呈色した色の濃さを目視判定
して評価されていた。このような目視判定は簡便で迅速
であり、十分な価値を有するものである。しかし、目視
判定では、当然精度が極めて低く、しかも同一色調の色
濃度は色見本等がないと判定しにくく、また色見本との
比較自体が熟練を要する。喘息等の対策としての屋内ア
レルゲン検査等、被検出物の量をできるだけ定量的に把
握したい場合は、上記のような検出方法では十分に対応
できなかった。特に屋内アレルゲンの場合、例えば布団
中のダニの量を検査した場合、検査結果に応じて、掃除
機等でダニを回収する等の早急な対策が必要であり、よ
り定量的な検出法が求められていた。In this conventional immunochromatography method, the presence or absence of the target antigen, ie, the object to be detected, is determined by the presence or absence of coloration at the detection portion, and the amount is evaluated by visually determining the color depth of the color. Was. Such visual judgment is simple and quick, and is of sufficient value. However, in the visual determination, the accuracy is naturally extremely low, and it is difficult to determine that the color density of the same color tone does not have a color sample or the like, and the comparison itself with the color sample requires skill. When it is desired to grasp the amount of an object to be detected as quantitatively as possible, such as indoor allergen test as a measure against asthma, etc., the above-described detection method cannot sufficiently cope. Especially in the case of indoor allergens, for example, when testing the amount of mites in the futon, urgent measures such as collecting mites with a vacuum cleaner etc. are required according to the test results, and a more quantitative detection method is required. Had been.
【０００４】[0004]
【発明が解決しようとする課題】本発明の目的は、溶液
中に採取した屋内アレルゲン等の存在量を、一般の人で
も、より定量的に、且つ簡便迅速に評価できる免疫検査
体を提供することである。SUMMARY OF THE INVENTION An object of the present invention is to provide an immunological test body which enables even ordinary people to more quantitatively, simply and quickly evaluate the abundance of indoor allergens and the like collected in a solution. That is.
【０００５】[0005]
【課題を解決するための手段】本発明は、一方はクロマ
ト展開可能な基材に固定されており、他方は着色微粒子
に固定されてクロマト展開前方に溶解可能な形で基材に
保持されている、同一被検出物の異なるエピトープに結
合する２種類の抗体対を含んでなる免疫検査体におい
て、同一の被検出物に対する検出感度が段階的に異なる
複数の単位免疫検査体から構成される免疫検査体に関す
る。特に、本発明は、検出感度の異なる複数の単位免疫
検査体が１個のクロマト基材上にクロマト展開方向に対
して平行に並列配置して組み込まれた上記免疫検査体に
関する。According to the present invention, one is fixed to a chromatographically expandable base material, and the other is fixed to colored microparticles and held by the base material in a form that can be dissolved before chromatographic development. An immunoassay comprising two types of antibody pairs that bind to different epitopes of the same analyte, the immunoassay comprising a plurality of unit immunoassays having different detection sensitivities for the same analyte in a stepwise manner Regarding the test object. In particular, the present invention relates to the above-described immunoassay, in which a plurality of unit immunoassays having different detection sensitivities are incorporated in a single chromatographic substrate in parallel and parallel to the chromatographic development direction.
【０００６】詳しくは、本発明は、各単位免疫検査体間
の検出感度の違いが固定された抗体の量の違いによって
形成されている上記免疫検査体に関する。もう一つの態
様では、本発明は、各単位免疫検査体間の検出感度の違
いがクロマト基材の展開速度の違いによって形成されて
いる上記免疫検査体に関する。また、詳しくは、本発明
は、検出感度の異なる複数の単位免疫検査体の検出表示
が異なる色標識によってなされる上記免疫検査体に関す
る。もう一つの態様では、本発明は、検出感度の異なる
複数の単位免疫検査体の検出表示が異なる形状をもつ表
示窓によってなされる上記免疫検査体に関する。更に、
本発明は、検体採取部、抗原溶解液収納部および上記免
疫検査体を含んでなるアレルゲン検査キットに関する。More specifically, the present invention relates to the above-mentioned immunoassay, wherein the difference in detection sensitivity between each unit immunoassay is formed by the difference in the amount of the fixed antibody. In another aspect, the present invention relates to the above-mentioned immunoassay, wherein the difference in detection sensitivity between each unit immunoassay is formed by the difference in the developing speed of the chromatographic substrate. More specifically, the present invention relates to the above-mentioned immunoassay, in which the detection display of a plurality of unit immunoassays having different detection sensitivities is made by different color labels. In another aspect, the present invention relates to the above-mentioned immunoassay, wherein detection displays of a plurality of unit immunoassays having different detection sensitivities are made by display windows having different shapes. Furthermore,
The present invention relates to an allergen test kit including a sample collection section, an antigen solution storage section, and the above-described immunological test body.
【０００７】[0007]
【発明の実施の形態】本発明では、検出感度の異なる複
数の単位免疫検査体が並列配置して１個の検査体として
組み込まれる。本発明において、並列配置とは、クロマ
ト展開可能なクロマト基材上に組み込まれた１対の標識
抗体と固定抗体とからなる抗体対を含んでなり特定抗原
を検出することのできる単位免疫検査体をそれぞれクロ
マト展開方向に対して平行に複数個並列配置して１個の
検査体として組み立てられた免疫検査体であって、個々
の単位免疫検査体はその一端である浸漬部が被検出体抗
原を溶解した溶液中に同時に浸漬されて、同時平行して
被検出体をクロマト展開できるようになっていることを
意味する。この場合、各単位免疫検査体は別のレーンを
形成していることになる。並列配置の方法は、図１に示
すように、すべての免疫検査体が１平面上に配置されて
もよいし（図１の（ａ））、三角柱、四角柱あるいは五
角柱のように立体的に組み立てられてもよい（図１の
（ｂ））。DESCRIPTION OF THE PREFERRED EMBODIMENTS In the present invention, a plurality of unit immunoassays having different detection sensitivities are arranged in parallel and incorporated as one examination body. In the present invention, the parallel arrangement means a unit immunoassay which comprises an antibody pair consisting of a pair of a labeled antibody and an immobilized antibody incorporated on a chromatographically expandable chromatographic substrate and can detect a specific antigen. Are arranged in parallel with each other in parallel to the direction of chromatographic development, and are assembled as one test body. In each unit immunological test body, the immersion part at one end is an antigen to be detected. Is simultaneously immersed in a solution in which is dissolved, so that the object to be detected can be developed in parallel at the same time. In this case, each unit immunoassay forms another lane. In the parallel arrangement method, as shown in FIG. 1, all the immunological test objects may be arranged on one plane (FIG. 1 (a)), or may be three-dimensional such as a triangular prism, a quadrangular prism or a pentagonal prism. (FIG. 1B).
【０００８】本発明の免疫検査体は、上記のように複数
個並列配置されて組み立てられた各単位免疫検査体が異
なる検出感度を持っているところに特徴がある。即ち、
免疫検査体はＮ個の単位免疫検査体Ａi（ｉ＝１，２，
・・・・・・，Ｎ）からなり、それぞれの検出感度ｓi
（ｉ＝１，２，・・・・・・，Ｎ）はｓ1＞ｓ2＞ｓ3＞
・・・・・＞ｓNとなっている。抗原量が少ない場合は
最も感度の高い単位免疫検査体Ａ1だけが抗原の存在を
感知することができ、Ａ1のみに、検出された抗原量に
応じた濃度で色表示される。Ａ1の感度限界を越えた量
の抗原が存在するとＡ1の検出部は飽和されてその色は
最高の濃度を示し、それ以上の抗原が到達してもＡ1は
同じ色濃度を保つか、またはプロゾーン現象により感度
が低下するだけであり、Ａ1だけではもはや濃度の違い
を表すことはできなくなる。しかし検査体には同時によ
り検出感度の低い単位免疫検査体Ａ2が組み込まれてい
るため、抗原濃度が高くなると今度はＡ2の方に色が表
示されるようになる。更に抗原の量が多くなるにつれて
より感度の低い単位免疫検査体の検出部に色表示が現れ
るようになる。このように検出感度の異なる複数の単位
免疫検査体を用いて同時にイムノクロマト試験を行うこ
とにより、存在する抗原の存在量範囲を容易に把握する
ことができる。The immunoassay of the present invention is characterized in that a plurality of unit immunoassays arranged and assembled in parallel as described above have different detection sensitivities. That is,
The immunity test object is composed of N unit immunity test objects A i (i = 1, 2, 2
.., N), and each detection sensitivity s i
(I = 1, 2,..., N) is s 1 > s 2 > s 3 >
...> S N. When the amount of antigen is small, only the unit immunoassay A 1 having the highest sensitivity can detect the presence of the antigen, and only A 1 is displayed in color at a concentration corresponding to the detected amount of antigen. Detection of A 1 when the amount of antigen that exceeds the limit of sensitivity A 1 is present the color is saturated in the highest concentration, or the A 1 be reached more antigen keep the same color density or the prozone phenomenon is only sensitivity is lowered, only a 1 will not be able anymore representing the difference in concentration. However since the built-in low unit immunoassay element A 2 detection sensitivity than simultaneously to the test body, this time when the antigen concentration increases will be displayed color towards the A 2. Further, as the amount of the antigen increases, a color display appears on the detection section of the unit immunoassay with lower sensitivity. By simultaneously performing an immunochromatographic test using a plurality of unit immunoassays having different detection sensitivities as described above, the range of the amount of the present antigen can be easily grasped.
【０００９】検出感度の異なる単位免疫検査体は次のよ
うにして作製することができる。一つの方法は、各単位
免疫検査体において同一の組み合わせの抗体対を用いる
が、単位免疫検査体ごとに固定抗体の量を変えることに
よる。イムノクロマト法においては、テストゾーンの固
定抗体に捕捉された抗原−標識抗体複合体の量が多いほ
どテストゾーンの色濃度が高くなる。したがって、検体
の抗原濃度が同程度である場合、固定抗体量が少ないと
呈色が低く、低感度となる。固定抗体量が多いと呈色が
強く、したがって固定抗体量を増加させた場合、検体中
の抗原濃度が低い場合でも呈色の目視認識が可能とな
る。[0009] Unit immunoassays having different detection sensitivities can be prepared as follows. One method uses the same combination of antibody pairs in each unit immunoassay, but by varying the amount of fixed antibody for each unit immunoassay. In the immunochromatography method, the larger the amount of the antigen-labeled antibody complex captured by the immobilized antibody in the test zone, the higher the color density in the test zone. Therefore, in the case where the antigen concentration of the specimen is almost the same, if the amount of the immobilized antibody is small, the coloration is low and the sensitivity is low. When the amount of the immobilized antibody is large, the color is strong. Therefore, when the amount of the immobilized antibody is increased, the color can be visually recognized even when the antigen concentration in the sample is low.
【００１０】もう一つの方法は、各単位免疫検査体にお
いて同一の組み合わせの抗体対を用いるが、それぞれで
異なる展開速度のクロマト基材を用いるものである。展
開液とともに同じ濃度で被検体抗原が運ばれて来ても、
展開速度が速い単位免疫検査体では標識抗体と結合する
ことなく下流へ通過する抗原量が増える。それに応じて
検出部で固定抗体により検出されるべき抗原の数が減少
する。即ち、展開速度の速い単位免疫検査体では検出感
度が低下する。したがって抗原量が少ない、即ち展開液
中の抗原濃度が低い場合は、展開速度の低い単位免疫検
査体では抗原の存在が表示されるが、展開速度がより高
い単位免疫検査体ではなお抗原は検出されない。抗原濃
度がより高くなるに連れてより展開速度の高い単位免疫
検査体でも固定抗体のところに到達する標識抗体付加抗
原の絶対量が増えるため抗原の存在が表示されるように
なる。したがってより展開速度の大きい単位免疫検査体
に表示が現れれば、その単位免疫検査体の濃度範囲に相
当した抗原が存在することを示していることになる。Another method is to use the same combination of antibody pairs in each unit immunoassay, but to use different chromatographic substrates for each. Even if the analyte antigen is carried at the same concentration with the developing solution,
In a unit immunoassay with a fast developing speed, the amount of antigen that passes downstream without binding to the labeled antibody increases. Accordingly, the number of antigens to be detected by the immobilized antibody in the detection section is reduced. That is, the detection sensitivity of a unit immunoassay having a high developing speed is reduced. Therefore, when the amount of antigen is small, that is, when the antigen concentration in the developing solution is low, the presence of the antigen is indicated in the unit immunoassay with a low developing speed, but the antigen is still detected in the unit immunoassay with a higher developing speed. Not done. As the antigen concentration becomes higher, even in a unit immunoassay having a higher developing speed, the absolute amount of the labeled antibody-added antigen reaching the fixed antibody increases, so that the presence of the antigen is displayed. Therefore, if a display appears on a unit immunoassay with a higher developing speed, it indicates that an antigen corresponding to the concentration range of the unit immunoassay exists.
【００１１】各感度の単位免疫検査体に於ける抗原の検
出表示は、それぞれの単位免疫検査体でレーンが異なる
ため、必ずしも区別する必要はないが、表示をより明瞭
にするために、検出感度の異なる単位免疫検査体ごとに
表示法を違えるのが好ましい。ひとつの表示方法は、感
度の異なる単位免疫検査体、即ちクロマトレーンごとに
抗体に取り付ける標識の色を変える方法である。例え
ば、感度の異なる３種類の単位免疫検査体を使用する場
合は、青、黄、赤の標識色を用いる方法である。これを
抗原濃度の許容量と関連付けて、青を許容できる濃度、
赤を警戒濃度として感度水準を設定することができる。
図２はこのような免疫検査体の例を示している。もう一
つの表示方法は、検出部の表示窓の形を変える方法であ
る。例えば、高感度の単位免疫検査体の表示窓は円形、
中感度の単位免疫検査体の表示窓は三角形、低感度の単
位免疫検査体の表示窓はバツ印（×）とする方法であ
る。この場合には円形表示で表示できる濃度を許容濃
度、バツ印で表示できる濃度を警戒濃度と関連付けて設
定することができる。図３はこのような免疫検査体の例
を示している。[0011] The detection of the antigen in the unit immunoassay of each sensitivity is not necessarily distinguished because the lanes are different in each unit immunoassay, but it is not always necessary to distinguish between them. It is preferable to use a different display method for each of the different unit immunoassays. One display method is to change the color of a label attached to an antibody for each unit immunoassay having a different sensitivity, that is, for each chromatane. For example, when three types of unit immunoassays having different sensitivities are used, a method using blue, yellow, and red marker colors is used. This is related to the allowable amount of antigen concentration, and the concentration that can tolerate blue,
The sensitivity level can be set using red as the alert density.
FIG. 2 shows an example of such an immunological test object. Another display method is a method of changing the shape of the display window of the detection unit. For example, the display window of a highly sensitive unit immunoassay is circular,
In this method, the display window of the medium-sensitivity unit immunoassay is triangular, and the display window of the low-sensitivity unit immunoassay is crossed (x). In this case, the density that can be displayed in a circular display can be set in association with the allowable density, and the density that can be displayed in a cross can be set in association with the alert density. FIG. 3 shows an example of such an immunological test object.
【００１２】また上記表示方法と関連させて、それぞれ
の感度の単位免疫検査体に表示が現れた場合の濃度範囲
をそれぞれの単位免疫検査体に記載しておいてもよい
し、濃度範囲を表示する代わりに、「注意。掃除機をか
けて下さい」、「要注意。掃除機をかけて下さい」等の
注意レベルを記載しておいてもよい。In connection with the above display method, the concentration range when a display appears on the unit immunoassay of each sensitivity may be described in each unit immunoassay, or the concentration range may be displayed. Instead, a caution level such as “Caution. Vacuum cleaner” or “Needs attention. Vacuum cleaner” may be described.
【００１３】本発明で使用することのできる各免疫検査
体は、図４に示すように、クロマト展開可能な基材上
に、試料溶液浸漬部６、標識抗体保持部３、検出部４お
よび試験終了確認部５が設けられている。As shown in FIG. 4, each immunoassay that can be used in the present invention is composed of a sample solution immersion section 6, a labeled antibody holding section 3, a detection section 4, and a test section on a chromatographically expandable substrate. An end confirmation unit 5 is provided.
【００１４】[0014]
【実施例】以下、実施例により、具体的に説明する。実施例 １ （１）免疫検査体の作製 １）標識抗体の調製 着色したポリスチレンラテックス（バングス・ラボラト
リーズ（Bangs Laboratories）社製）中に抗体（抗Ｄｅ
ｒ ｐ１抗体）を混合し、インキュベートして、抗体を
ポリマー粒子に固定した。ラテックス粒子として、赤、
青および黄色の３種類のものを選ぶことにより、３種類
の標識抗体を得た。The present invention will be specifically described below with reference to examples. Example 1 (1) Preparation of Immunoassay 1) Preparation of Labeled Antibody Antibody (anti-De) was added to a colored polystyrene latex (manufactured by Bangs Laboratories).
rp1 antibody) was mixed and incubated to immobilize the antibody on the polymer particles. Red, as latex particles
Three kinds of labeled antibodies were obtained by selecting three kinds of blue and yellow.
【００１５】２）免疫検査体の作製 １ｃｍ×１０ｃｍ（厚さ０.７ｍｍ）のグラスフィルタ
ー（ワットマン社製）をクロマト基材として準備した。
これに３つのレーンを形成し、それぞれの下流に当たる
部分に抗原検出用固定抗体として働く抗ダニアレルゲン
抗体液（抗ダニアレルゲン抗体を生理食塩水中に溶解し
たもの）１μｌをピペットマンで滴下し、次に、このグ
ラスフィルター上の各レーンの、更に下流部に、標識抗
体と反応して試験終了確認用として働く抗ＩｇＧ抗体液
１μｌをピペットマンで滴下した。ただし、抗アレルゲ
ン抗体液の濃度は３種類の濃度のものを使用し、３本の
レーンのうち左側のレーンには最も濃度の高い１０ｍｇ
／ｍｌのもの、中央のレーンには次に高い濃度１ｍｇ／
ｍｌのもの、右側のレーンには最も低い濃度０.１ｍｇ
／ｍｌの抗体溶解液を滴下して固定した。このグラスフ
ィルターを５０℃で１分間乾燥して２種類の抗体をグラ
スフィルターに固定した。次に、上で調製した３種類の
標識抗体を、このグラスフィルターの上流部分に、各レ
ーンごとに異なる色のものを１５μｌずつ滴下した。即
ち左側のレーンＡ1には青色、中央のレーンＡ2には黄
色、右側のレーンＡ3には赤色の標識抗体を滴下した。
標識抗体を滴下したグラスフィルターを５０℃で５分間
乾燥して標識抗体をグラスフィルター上に保持した。上
記において、標識抗体保持部、抗アレルゲン抗体固定部
および抗ＩｇＧ抗体固定部の位置はそれぞれ、グラスフ
ィルターの検体供与側の端部から３ｃｍ、７ｃｍおよび
８ｃｍとした。2) Preparation of immunological test body A glass filter (manufactured by Whatman) having a size of 1 cm × 10 cm (0.7 mm in thickness) was prepared as a chromatographic substrate.
Three lanes are formed in this, and 1 μl of an anti-mite allergen antibody solution (a solution of the anti-mite allergen antibody dissolved in physiological saline) serving as a fixed antibody for antigen detection is dropped by a pipetman onto portions downstream of each of the lanes. Then, 1 μl of an anti-IgG antibody solution that reacts with the labeled antibody and serves as a confirmation of the end of the test was dropped by a pipetteman further downstream of each lane on the glass filter. However, the concentration of the anti-allergen antibody solution used was one of three concentrations.
/ Ml, the next highest concentration of 1 mg /
ml, the lowest concentration is 0.1 mg in the right lane.
/ Ml of the antibody solution was dropped and fixed. The glass filter was dried at 50 ° C. for 1 minute to fix two kinds of antibodies on the glass filter. Next, 15 μl of the above-prepared three types of labeled antibodies having different colors for each lane were dropped on the upstream portion of the glass filter. That is, the lane A 1 of the left blue, the center of the lane A 2 yellow, the right lane A 3 was added dropwise a red-labeled antibody.
The glass filter to which the labeled antibody had been dropped was dried at 50 ° C. for 5 minutes to hold the labeled antibody on the glass filter. In the above description, the positions of the labeled antibody holding section, the anti-allergen antibody fixing section and the anti-IgG antibody fixing section were respectively 3 cm, 7 cm and 8 cm from the end of the glass filter on the sample donor side.
【００１６】（２）免疫検査体の使用例 １ｍ2の布団に１分間掃除機をかけて採取したゴミを２
０ｍｌの抗原溶解液に混合し、濾過して固形分を除去し
て検体液を調製した。この検体液に、上記で作製した免
疫検査体の最上流部である浸漬部を浸漬し、免疫検査体
を垂直に保持した状態で５秒間静置した。５秒後、左側
の青色標識抗体のレーンの検出部および試験終了確認部
は濃い青色に着色し、また中央レーンの検出部は薄い黄
色に着色していた。なお中央および右側レーンの試験終
了確認部はそれぞれ濃い黄色および赤色に着色してい
た。[0016] (2) waste that was collected over a period of 1 minute vacuum cleaner to the futon of use example 1m 2 of immunoassay body 2
The mixture was mixed with 0 ml of the antigen solution and filtered to remove solids, thereby preparing a sample solution. The immersion part, which is the uppermost stream of the immunological test specimen prepared above, was immersed in this sample liquid, and left standing for 5 seconds with the immunological test specimen held vertically. Five seconds later, the detection part of the left blue labeled antibody lane and the test completion confirmation part were colored dark blue, and the detection part of the center lane was pale yellow. Note that the test completion confirmation sections in the center and right lanes were colored deep yellow and red, respectively.
【００１７】実施例 ２ （１）単位免疫検査体の作製 １）標識抗体の調製 実施例１と同様にして赤色の標識抗体を調製した。 ２）単位免疫検査体の作製 構成する繊維の繊度および目付量のことなる３種類のグ
ラスファイバーを準備し、それぞれを１ｃｍ×１０ｃｍ
（厚さ０.７ｍｍ）に裁断した。それぞれにグラスファ
イバーの下流に当たる同じ部分に抗原検出用固定抗体と
して働く抗ダニアレルゲン抗体液（抗ダニアレルゲン抗
体を生理食塩水中に濃度１ｍｇ／ｍｌで溶解したもの）
１μｌをピペットマンで滴下し、次に、この各グラスフ
ィルター上の更に下流部の同じ位置に、標識抗体と反応
して試験終了確認用として働く抗ＩｇＧ抗体液（抗Ｉｇ
Ｇ抗体を生理食塩水中に１ｍｇ／ｍｌの濃度で溶解した
もの）１μｌをピペットマンで滴下した。このグラスフ
ィルターを５０℃で１分間乾燥して２種類の抗体をグラ
スフィルターに固定した。 Example 2 (1) Preparation of unit immunoassay 1) Preparation of labeled antibody A red labeled antibody was prepared in the same manner as in Example 1. 2) Preparation of unit immunoassay Three kinds of glass fibers having different fineness and basis weight of the constituent fibers were prepared, and each of them was 1 cm × 10 cm.
(Thickness: 0.7 mm). Anti-mite allergen antibody solution (anti-mite allergen antibody dissolved in physiological saline at a concentration of 1 mg / ml) that acts as a fixed antibody for antigen detection in the same portion downstream of the glass fiber.
1 μl was dropped with a pipetman, and then, at the same position further downstream on each glass filter, an anti-IgG antibody solution (anti-Ig
1 μl of G antibody dissolved in physiological saline at a concentration of 1 mg / ml) was added dropwise using a pipetman. The glass filter was dried at 50 ° C. for 1 minute to fix two kinds of antibodies on the glass filter.
【００１８】次に、上で調製した標識抗体を、このグラ
スフィルターの上流部分の同じ位置に１５μｌずつ滴下
し、５０℃で５分間乾燥して標識抗体をグラスフィルタ
ー上に保持した。次に、グラスファイバー上の固定抗体
を植え付けた部分をポリエステル製の薄い不透明シート
で被覆した。抗ＩｇＧ抗体を固定した部分には抗ＩｇＧ
抗体の変色が観察できるようにライン状の切り抜きを入
れた。また抗ダニアレルゲン抗体を固定した部分には図
形の切り抜きを入れた。ただし、切り抜きの形は、グラ
スファイバーの種類によって違え、目付量が最大のもの
は円形、中位のものは三角、目付量が最小のものはバツ
印とした。こうして３種類の単位免疫検査体Ａ1、Ａ2、
Ａ3を得た。最後に３種類の単位免疫検査体を平行した
３つのレーンを形成するように張り合わせて免疫検査体
を得た。Next, the labeled antibody prepared above was dropped at the same position on the upstream part of the glass filter in 15 μl portions, and dried at 50 ° C. for 5 minutes to hold the labeled antibody on the glass filter. Next, the portion of the glass fiber where the immobilized antibody was implanted was covered with a thin opaque sheet made of polyester. The portion where the anti-IgG antibody is immobilized is
A line-shaped cutout was made so that discoloration of the antibody could be observed. In addition, a cutout of a figure was made in a portion where the anti-mite allergen antibody was fixed. However, the shape of the cutout was different depending on the type of glass fiber. The shape with the largest weight was circular, the shape with a medium weight was triangular, and the shape with the smallest weight was crossed. Thus, three types of unit immunoassays A 1 , A 2 ,
To give the A 3. Finally, three types of unit immunoassays were stuck together to form three parallel lanes to obtain an immunoassay.
【００１９】[0019]
【発明の効果】本発明の免疫検査体は、抗原抗体反応に
より被検体を検出する従来の免疫検査体の限界であった
定量性を改良したものであり、存在する被検体の量を範
囲として検出することができる。したがってこの免疫検
査体を用いてダニ等のアレルゲンを測定することによ
り、アレルゲンの存在量を把握することができ、より適
切な対応を選ぶことが可能となる。The immunoassay of the present invention is an improvement of the quantitative property, which was the limit of the conventional immunoassay for detecting a subject by an antigen-antibody reaction, and the range of the amount of the existing analyte is limited. Can be detected. Therefore, by measuring allergens such as mites using this immune test specimen, the amount of allergens present can be ascertained, and more appropriate measures can be selected.
【図１】 本発明の免疫検査体の示す模式図。（ａ）各
単位免疫検査体を平面状に配置した免疫検査体、（ｂ）
各単位免疫検査体を立体的に配置した免疫検査体の１
例。FIG. 1 is a schematic diagram showing an immunological test body of the present invention. (A) an immunoassay body in which each unit immunoassay is arranged in a plane, (b)
One of the immunoassays in which each unit immunoassay is arranged three-dimensionally
Example.
【図２】 検出感度を色の違いによって表示する免疫検
査体の例。FIG. 2 shows an example of an immunological test object displaying detection sensitivity by different colors.
【図３】 検出感度をパターンの違いによって表示する
免疫検査体の例。FIG. 3 is an example of an immunological test object displaying detection sensitivity by a difference in pattern.
【図４】 免疫検査体の構成を示す模式図。FIG. 4 is a schematic diagram showing a configuration of an immunological test object.
１：免疫検査体、 ２：単位免疫検査体、
３：標識抗体保持部、 ４：検出表示部、５：
試験終了確認部、 ６：浸漬部、７：吸液材
８：基材、９：固定抗体部カバー。1: immunological test, 2: unit immunological test,
3: Labeled antibody holding part, 4: Detection display part, 5:
Test completion confirmation part, 6: immersion part, 7: liquid absorbing material
8: base material, 9: fixed antibody part cover.
Claims (9)
れており、他方は着色微粒子に固定されてクロマト展開
前方に溶解可能な形で基材に保持されている、同一被検
出物の異なるエピトープに結合する２種類の抗体対を含
んでなる免疫検査体において、同一の被検出物に対する
検出感度が段階的に異なる複数の単位免疫検査体から構
成される免疫検査体。1. One is fixed to a chromatographically developable base material, and the other is fixed to colored fine particles and held by the base material in a form dissolvable in front of chromatographic development. An immunoassay comprising two types of antibody pairs that bind to an epitope, wherein the immunoassay is composed of a plurality of unit immunoassays having different detection sensitivities to the same analyte in a stepwise manner.
が１個のクロマト基材上にクロマト展開方向に対して平
行に並列配置して組み込まれた請求項１記載の免疫検査
体。2. The immunoassay according to claim 1, wherein a plurality of unit immunoassays having different detection sensitivities are arranged in parallel on a single chromatographic substrate in parallel with the direction of chromatographic development.
固定された抗体の量の違いによって形成されている請求
項１または２に記載の免疫検査体。3. The immunoassay according to claim 1, wherein a difference in detection sensitivity between the unit immunoassays is formed by a difference in the amount of the immobilized antibody.
クロマト基材の展開速度の違いによって形成されている
請求項１または２記載の免疫検査体。4. The immunoassay according to claim 1, wherein a difference in detection sensitivity between the unit immunoassays is formed by a difference in a developing speed of the chromatographic substrate.
の検出表示が異なる色標識によってなされる請求項１〜
４のいずれかひとつに記載の免疫検査体。5. The detection display of a plurality of unit immunoassays having different detection sensitivities is performed by different color labels.
4. The immunological test body according to any one of 4 above.
スによってなされる請求項５記載の免疫検査体。6. The immunoassay according to claim 5, wherein the different colored markers are formed by colored latexes of different colors.
の検出表示が異なる形状をもつ表示窓によってなされる
請求項１〜４のいずれかひとつに記載の免疫検査体。7. The immunoassay according to claim 1, wherein the detection display of a plurality of unit immunoassays having different detection sensitivities is performed by display windows having different shapes.
色ラテックスによってなされる請求項６記載の免疫検査
体。8. The immunoassay according to claim 6, wherein the display window is labeled with gold colloid or colored latex.
求項１〜９のいずれかひとつに記載の免疫検査体を含ん
でなるアレルゲン検査キット。9. An allergen test kit comprising a sample collection section, an antigen solution storage section, and the immunological test body according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP13022298A JPH11326326A (en) | 1998-05-13 | 1998-05-13 | Immunity inspection body |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP13022298A JPH11326326A (en) | 1998-05-13 | 1998-05-13 | Immunity inspection body |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH11326326A true JPH11326326A (en) | 1999-11-26 |
Family
ID=15029018
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP13022298A Pending JPH11326326A (en) | 1998-05-13 | 1998-05-13 | Immunity inspection body |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH11326326A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001235471A (en) * | 2000-02-23 | 2001-08-31 | Nippon Chemiphar Co Ltd | Method using porous filter for measuring bioactive sample substance |
| WO2003029822A1 (en) * | 2001-09-28 | 2003-04-10 | Matsushita Electric Industrial Co., Ltd. | Specific binding analyzer and method of analyzing specific binding |
| WO2006011249A1 (en) * | 2004-07-28 | 2006-02-02 | Nippon Gene Co., Ltd | Analytical apparatus and method of analysis |
| JP2013113711A (en) * | 2011-11-29 | 2013-06-10 | Kddi Corp | Biosensor and method for measuring test solution |
| WO2017078170A1 (en) * | 2015-11-05 | 2017-05-11 | 武 永安 | Monoclonal antibody specific to ck19, hybridoma for producing monoclonal antibody, kit for detecting cancer, method for detecting cancer, and method for determining cancer metastasis |
| JP2019514009A (en) * | 2016-04-13 | 2019-05-30 | ノヴァメド リミテッド | One-step cardiac examination device |
-
1998
- 1998-05-13 JP JP13022298A patent/JPH11326326A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001235471A (en) * | 2000-02-23 | 2001-08-31 | Nippon Chemiphar Co Ltd | Method using porous filter for measuring bioactive sample substance |
| WO2003029822A1 (en) * | 2001-09-28 | 2003-04-10 | Matsushita Electric Industrial Co., Ltd. | Specific binding analyzer and method of analyzing specific binding |
| WO2006011249A1 (en) * | 2004-07-28 | 2006-02-02 | Nippon Gene Co., Ltd | Analytical apparatus and method of analysis |
| JP2006038700A (en) * | 2004-07-28 | 2006-02-09 | Nippon Gene Co Ltd | Analyzer and analysis method |
| JP2013113711A (en) * | 2011-11-29 | 2013-06-10 | Kddi Corp | Biosensor and method for measuring test solution |
| WO2017078170A1 (en) * | 2015-11-05 | 2017-05-11 | 武 永安 | Monoclonal antibody specific to ck19, hybridoma for producing monoclonal antibody, kit for detecting cancer, method for detecting cancer, and method for determining cancer metastasis |
| JP2017088519A (en) * | 2015-11-05 | 2017-05-25 | 武 永安 | Ck19 specific monoclonal antibodies and hybridoma producing the same, cancer detection kits, cancer detection methods as well as determination methods of cancer metastasis |
| JP2019514009A (en) * | 2016-04-13 | 2019-05-30 | ノヴァメド リミテッド | One-step cardiac examination device |
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