JPH02254364A - Plate for measuring immunity of biological/chemical luminescent enzyme - Google Patents
Plate for measuring immunity of biological/chemical luminescent enzymeInfo
- Publication number
- JPH02254364A JPH02254364A JP1077529A JP7752989A JPH02254364A JP H02254364 A JPH02254364 A JP H02254364A JP 1077529 A JP1077529 A JP 1077529A JP 7752989 A JP7752989 A JP 7752989A JP H02254364 A JPH02254364 A JP H02254364A
- Authority
- JP
- Japan
- Prior art keywords
- sample
- light
- sample hole
- plate
- holes
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000000126 substance Substances 0.000 title abstract description 7
- 102000004190 Enzymes Human genes 0.000 title description 18
- 108090000790 Enzymes Proteins 0.000 title description 18
- 230000036039 immunity Effects 0.000 title 1
- 229920003002 synthetic resin Polymers 0.000 claims abstract description 10
- 239000000057 synthetic resin Substances 0.000 claims abstract description 10
- 229910052751 metal Inorganic materials 0.000 claims abstract description 8
- 239000002184 metal Substances 0.000 claims abstract description 8
- 239000000843 powder Substances 0.000 claims abstract description 8
- 238000012767 chemiluminescent enzyme immunoassay Methods 0.000 claims description 7
- 238000004166 bioassay Methods 0.000 claims description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 3
- 239000003973 paint Substances 0.000 claims description 3
- 238000005259 measurement Methods 0.000 abstract description 30
- 239000011248 coating agent Substances 0.000 abstract description 12
- 238000004020 luminiscence type Methods 0.000 abstract description 6
- 238000002796 luminescence method Methods 0.000 abstract description 4
- 229910052782 aluminium Inorganic materials 0.000 abstract description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 abstract description 2
- 239000012295 chemical reaction liquid Substances 0.000 abstract description 2
- 238000006243 chemical reaction Methods 0.000 description 18
- 238000000034 method Methods 0.000 description 12
- 239000000427 antigen Substances 0.000 description 9
- 102000036639 antigens Human genes 0.000 description 9
- 108091007433 antigens Proteins 0.000 description 9
- 238000003018 immunoassay Methods 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 6
- 230000035945 sensitivity Effects 0.000 description 5
- 102000003992 Peroxidases Human genes 0.000 description 4
- 238000002795 fluorescence method Methods 0.000 description 4
- 108040007629 peroxidase activity proteins Proteins 0.000 description 4
- 238000004737 colorimetric analysis Methods 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 238000005415 bioluminescence Methods 0.000 description 2
- 230000029918 bioluminescence Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 239000007800 oxidant agent Substances 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
- G01N21/0303—Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
- G01N2021/0378—Shapes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6463—Optics
- G01N2021/6469—Cavity, e.g. ellipsoid
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6482—Sample cells, cuvettes
Landscapes
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Optical Measuring Cells (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は近年医学、薬学又は生化学等の分野において利
用されている酵素免疫測定法における生物発光、化学発
光を用いた抗原検出方法に最適な生物、化学発光酵素免
疫測定用プレートに関するものである。Detailed Description of the Invention (Field of Industrial Application) The present invention is most suitable for antigen detection methods using bioluminescence and chemiluminescence in enzyme immunoassay methods that have been used in the fields of medicine, pharmacy, biochemistry, etc. in recent years. This invention relates to plates for immunoassay of biological organisms and chemiluminescent enzymes.
（従来の技術）
酵素免疫測定法（エンザイム・イムノアッセイ）は、予
め試料台である酵素免疫測定用プレートの有底試料孔に
、被測定対象物に対する特異抗体を結合させておき（抗
体の固相化）、これに被測定対象物である抗原を含む血
液等の試料液を加えて免疫反応（抗原抗体反応）を行な
わせ、抗原を固相化抗体に結合させる。次に、未反応物
を洗浄して除去した後、固相化抗体に結合した抗原に、
さらに酵素標識抗体を加えて結合させ、この抗原に結合
した酵素標識抗体の量を測定することにより、被測定対
象物である抗原の量を求める方法である。(Prior art) In enzyme immunoassay, a specific antibody against the object to be measured is bound in advance to the bottomed sample hole of an enzyme immunoassay plate (sample stage). A sample liquid such as blood containing an antigen to be measured is added to this to cause an immune reaction (antigen-antibody reaction), and the antigen is bound to the immobilized antibody. Next, after washing and removing unreacted substances, the antigen bound to the immobilized antibody is
In this method, an enzyme-labeled antibody is further added and bound to the antigen, and the amount of the enzyme-labeled antibody bound to the antigen is measured, thereby determining the amount of the antigen to be measured.
酵素標識抗体の検出方法としては、通常、比色法あるい
は蛍光法が用いられている。比色法は、基質を加えて反
応させ、その吸光度を測定する方法である。蛍光法は、
基質に蛍光物質を用い、反応させた後、光で励起して蛍
光を起こし、その蛍光強度を測定する方法である。前者
は、操作の簡便さから従来よく用いられているが、感度
は低い。As a method for detecting enzyme-labeled antibodies, a colorimetric method or a fluorescence method is usually used. The colorimetric method is a method in which a substrate is added and reacted, and its absorbance is measured. Fluorescence method is
This method uses a fluorescent substance as a substrate, causes a reaction, and then excites it with light to cause fluorescence, and then measures the intensity of the fluorescence. The former method has been commonly used due to its ease of operation, but its sensitivity is low.
一方、後者は、前者の１００〜１，０００倍の感度を示
すものの、励起光を必要とするために、絶えず被測定試
料以外の物質の干渉を招き、実用されていないのが現状
である。これらの理由から、高感度化の方法として放射
免疫測定法が使用されてきたが、この方法は、標識物質
として放射性同位元素を使用するために取り扱いが限定
され、そのうえ測定者への安全上の問題もあるため、こ
れに変わる超高感度測定法が求められてきた。On the other hand, although the latter exhibits a sensitivity 100 to 1,000 times higher than the former, it requires excitation light, which constantly causes interference with substances other than the sample to be measured, and is currently not in practical use. For these reasons, radioimmunoassay has been used as a method to increase sensitivity, but this method uses a radioisotope as a labeling substance, which limits its handling, and it also poses safety concerns for the person performing the measurement. Due to these problems, an alternative, ultra-sensitive measurement method has been sought.
近年、これらの問題を解決する方法として、生物あるい
は化学発光酵素免疫測定法が注目を浴びている。その大
きな理由は、発光法は、蛍光法のように励起光を必要と
せず、適当な酸化剤を添加すれば、抗原に特異的に結合
した酵素（酵素標識抗体）の触媒によって、被測定試料
そのものがきわめて特異的に発光するためである。また
、蛍光法に比べて反応によって生ずる発光が微弱である
という問題があったが、光子計数法を採用することによ
り、高い感度を得ることができるようになった。In recent years, biological or chemiluminescent enzyme immunoassay methods have attracted attention as methods for solving these problems. The main reason for this is that the luminescence method does not require excitation light like the fluorescence method, and when an appropriate oxidizing agent is added, the sample to be measured is catalyzed by an enzyme (enzyme-labeled antibody) that specifically binds to the antigen. This is because the light itself emits light in a very specific manner. Furthermore, compared to the fluorescence method, there was a problem in that the light emitted by the reaction was weaker, but by adopting the photon counting method, it became possible to obtain high sensitivity.
また従来、このような酵素免疫測定法では、数多くの試
料を迅速にしかも正確に測定するために、反応容量の少
ない規則的に配設された多数の試料孔を有する酵素免疫
測定用プレートが使用されている。Conventionally, such enzyme immunoassay methods use enzyme immunoassay plates that have a large number of regularly arranged sample holes with a small reaction volume in order to measure a large number of samples quickly and accurately. has been done.
（発明が解決しようとする課題）
しかしながら、従来の酵素免疫測定用プレート（以下、
単に測定用プレートという。）は、主に光を透過させる
必要性のある比色法で使用されるものであることから、
第７図に示すように、透明な合成樹脂からなり、しかも
試料孔２の底が平底あるいはＶ底又はＵ底に形成されて
いる。そのため、これらの測定用プレートは、発光法に
用いることはできない。この理由は、第７図に示すよう
に、測定用プレートｌａが透明であると隣接する試料孔
２での発光と干渉したり、また、底が平底、■底等であ
ると受光部５への集光性が悪くなるため、このような従
来の測定用プレートｌａでは被測定試料孔２において試
料の発光量が正確に測定できないからである。(Problem to be solved by the invention) However, the conventional enzyme immunoassay plate (hereinafter referred to as
It is simply called a measurement plate. ) is mainly used in colorimetric methods that require light to pass through.
As shown in FIG. 7, it is made of transparent synthetic resin, and the bottom of the sample hole 2 is formed into a flat bottom, a V bottom, or a U bottom. Therefore, these measurement plates cannot be used in the luminescence method. The reason for this is that, as shown in FIG. 7, if the measuring plate la is transparent, it will interfere with the light emitted from the adjacent sample hole 2, and if the measuring plate la has a flat bottom, This is because such a conventional measurement plate la cannot accurately measure the amount of light emitted by the sample in the sample hole 2 because of the poor light focusing ability.
そこで、各試料孔２毎に１孔ずつ発光反応を行なわせ、
確実に受光して計測することも考えられるが、この方法
では、−度に多数の試料を測定することができず、迅速
性に欠けるという欠点がある。Therefore, a luminescent reaction was performed for each sample hole 2,
Although it is possible to reliably receive and measure light, this method has the disadvantage that it is not possible to measure a large number of samples at once, and it lacks speed.
本発明は斯かる事情に鑑みてなされたもので、集光性が
良く、しかも隣接する試料孔の発光の影響のない正確で
迅速な測定が行なえ、生物発光。The present invention has been developed in view of the above circumstances, and it has good light focusing ability, and can perform accurate and quick measurements without being affected by the luminescence of adjacent sample holes.
化学発光を用いた酵素免疫測定法に最適な測定用プレー
トを提供することを目的とする。The purpose of this invention is to provide an optimal measurement plate for enzyme immunoassay using chemiluminescence.
（課題を解決するための手段）
前記目的を達成するため、本発明は、多数の有底試料孔
を設けて、各試料孔の底を曲面で形成するとともに、各
試料孔の少なくとも側部及び底部に非透光性処理を施し
たものである。(Means for Solving the Problem) In order to achieve the above object, the present invention provides a large number of sample holes with a bottom, and forms the bottom of each sample hole with a curved surface, and at least the side and side portions of each sample hole. The bottom part has been treated to make it non-transparent.
前記試料孔の少なくとも側部及び底部は、その外面に金
属粉又は黒色塗料等をコーティングしたもの、金属粉末
又は炭素粉末等を混入した合成樹脂で形成したもの、あ
るいは着色合成樹脂で形成したものとすることができる
。At least the side and bottom portions of the sample hole may be coated with metal powder or black paint on the outer surface, made of synthetic resin mixed with metal powder or carbon powder, or made of colored synthetic resin. can do.
（作用）
各試料孔で得られた酵素標識抗体の生物発光又は化学発
光反応による光は、非透光性処理が施されて鏡面となり
、かつ、曲面で形成された試料孔の底部で反射して試料
孔の開口部方向に向かって進行し、該開口部近傍に配設
される受光部に集光される。また、試料孔の少なくとも
側部及び底部は、非透光性処理が施されているため、隣
接する試料孔からの光が遮光される。(Function) The light from the bioluminescence or chemiluminescence reaction of the enzyme-labeled antibody obtained in each sample hole is treated to make it non-transparent and becomes a mirror surface, and is reflected at the bottom of the sample hole, which is formed with a curved surface. The light travels toward the opening of the sample hole, and is focused on a light receiving section disposed near the opening. Furthermore, since at least the side and bottom portions of the sample hole are treated to make them non-transparent, light from adjacent sample holes is blocked.
（実施例） 次に、本発明の実施例を添付図面に従って説明する。(Example) Next, embodiments of the present invention will be described with reference to the accompanying drawings.
第３図は、本発明に係る測定用プレート１を示し、この
測定用プレートｌは透明な合成樹脂からなり、上面に規
則的に配列された多数の試料孔２を有している（ただし
、図では一部の試料孔２のみを示し、他は省略しである
。）。FIG. 3 shows a measurement plate 1 according to the present invention, which is made of transparent synthetic resin and has a large number of regularly arranged sample holes 2 on its upper surface (however, In the figure, only some of the sample holes 2 are shown, and the others are omitted.)
各試料孔２は、有底で竿１図に示すように、その底面は
曲面で形成されている。この底面の曲面形状としては、
放物面、二次曲面あるいは半楕円曲面等が好ましい。ま
た、各試料孔２の底外面。Each sample hole 2 has a bottom, and as shown in FIG. 1, the bottom surface is formed as a curved surface. The curved shape of this bottom surface is
A paraboloid, a quadratic curved surface, a semi-elliptic curved surface, etc. are preferable. Also, the bottom outer surface of each sample hole 2.
外側面及び測定用プレート１の裏面は、アルミニウム等
の金属粉を含むコーティング剤３でコーティングされて
いる。このコーティング剤３をコーティングする代わり
に黒色系塗料を塗布してもよい。The outer surface and the back surface of the measurement plate 1 are coated with a coating agent 3 containing metal powder such as aluminum. Instead of coating with this coating agent 3, a black paint may be applied.
第１図は、本実施例に係る測定用プレート１を用いて、
化学発光酵素免疫測定法により血液等の被測定試料中の
特定の抗原を分析する一過程を示す。４は、ベルオキシ
ターゼを標識した酵素標識抗体に、発光基質であるルミ
ノールと酸化剤である過酸化水素を加えた化学発光反応
液である。５は、この化学発光反応液４が発する光の量
を測定するルミノメータの受光部で、各試料孔２の開口
部近傍に位置している。FIG. 1 shows that using the measurement plate 1 according to this embodiment,
This figure shows a process of analyzing a specific antigen in a sample to be measured such as blood using chemiluminescent enzyme immunoassay. 4 is a chemiluminescent reaction solution prepared by adding luminol, which is a luminescent substrate, and hydrogen peroxide, which is an oxidizing agent, to an enzyme-labeled antibody labeled with peroxidase. Reference numeral 5 denotes a light receiving part of a luminometer that measures the amount of light emitted by the chemiluminescent reaction liquid 4, and is located near the opening of each sample hole 2.
化学発光反応液４が発する光は、四方に放射されるが、
試料孔２の底面がコーティング剤３により鏡面となって
おり、しかも曲面で形成されているため、この試料孔２
の底面で反射し、試料孔２の開口部に向かって進行して
受光部５に集光される。また、コーティング剤３により
、外部からの光、特に隣接する試料孔２からの光が遮光
される。The light emitted by the chemiluminescent reaction solution 4 is radiated in all directions,
The bottom surface of the sample hole 2 has a mirror surface due to the coating agent 3, and is also formed with a curved surface.
The light is reflected at the bottom surface of the sample hole 2, travels toward the opening of the sample hole 2, and is focused on the light receiving section 5. Furthermore, the coating agent 3 blocks light from the outside, particularly light from the adjacent sample hole 2 .
これにより、受光部５には、それが位置する被測定用試
料孔２からの化学発光による光のみが集光されるため、
発光量が正確に測定できる。また、隣接する試料孔２で
の化学発光の影響を受けないので、各試料孔２で一度に
化学発光させることができ、測定が迅速に行なえる。As a result, only light due to chemiluminescence from the sample hole 2 to be measured in which the light receiving part 5 is located is focused on the light receiving part 5.
The amount of light emitted can be measured accurately. Moreover, since it is not affected by chemiluminescence in adjacent sample holes 2, chemiluminescence can be caused in each sample hole 2 at once, and measurements can be performed quickly.
第２図は、本発明の他の実施例に係る測定用プレート１
０を示し、この測定用プレート１０は、予め金属粉末又
は炭素粉末等を混入した合成樹脂で形成して非透光性と
したものであり、コーティング剤を塗布していない以外
は第１図に示す測定用プレート１と同様であり、対応す
る部分には同一符号を付して説明を省略する。FIG. 2 shows a measuring plate 1 according to another embodiment of the present invention.
0, and this measurement plate 10 is made of synthetic resin mixed with metal powder or carbon powder in advance to make it non-transparent, and the measurement plate 10 is similar to that shown in FIG. 1 except that it is not coated with a coating agent. It is the same as the measurement plate 1 shown in FIG.
なお、第２図に示す金属粉末等混入の合成樹脂の代わり
に着色合成樹脂を用いてもよい。Note that a colored synthetic resin may be used instead of the synthetic resin mixed with metal powder etc. shown in FIG.
本発明者らは、本発明に係る測定用プレート１゜１０の
有効性を確認するために以下に示す実験を行なった。The present inventors conducted the following experiment to confirm the effectiveness of the measurement plate 1.10 according to the present invention.
実験１．外部光の影響
第４図に示すように、化学発光反応を行なわせてその発
光量を検出する１個の被測定試料孔２ａを特定し、該被
測定試料孔２ａの周囲に化学発光反応のみを行なわせる
反応試料孔２ｂを設ける。Experiment 1. Influence of external light As shown in Fig. 4, one sample hole 2a to be measured in which a chemiluminescent reaction is to be carried out and its luminescence amount is detected is specified, and only the chemiluminescent reaction is placed around the sample hole 2a to be measured. A reaction sample hole 2b is provided to allow the reaction to take place.
そして、反応試料孔２ｂの数を１個ずつ増加させて、被
測定試料孔２ａでの受光部５が受光する受光値すなわち
光電子パルス数を測定した。Then, the number of reaction sample holes 2b was increased one by one, and the light reception value, that is, the number of photoelectron pulses received by the light receiving section 5 in the sample hole 2a to be measured was measured.
第５図はこの実験結果を示し、Ａは本発明に係る測定用
プレートｌを用いたもの、Ｂは従来の測定用プレートを
用いたものである。ただし、従来の測定用プレートとし
ては、Ｕ底でコーティング剤を塗布していない透明のも
のを用いた。FIG. 5 shows the results of this experiment, where A is the result using the measurement plate 1 according to the present invention, and B is the result using the conventional measurement plate. However, as the conventional measurement plate, a transparent plate with a U bottom and no coating agent was used.
この実験１によれば、従来の測定プレートでは、反応試
料孔数の増加に伴い、被測定試料孔２ａでの受光値が増
大するため、周囲の反応試料孔２ｂでの化学発光の影響
を受けていることがわかる。According to this experiment 1, in the conventional measurement plate, as the number of reaction sample holes increases, the light reception value at the measurement sample hole 2a increases, so it is affected by chemiluminescence from the surrounding reaction sample holes 2b. You can see that
これに対し、本発明に係る測定用プレート１では、反応
試料孔数の増加にかかりらず被測定試料孔２ａでの受光
値は変化しないため、周囲の反応試料孔２ｂでの化学発
光の影響は全く受けていないことがわかる。On the other hand, in the measurement plate 1 according to the present invention, the light reception value in the measurement sample hole 2a does not change even though the number of reaction sample holes increases, so the influence of chemiluminescence in the surrounding reaction sample holes 2b It can be seen that it has not been received at all.
実験２．集光性
底形状がＵ底、■底、平底の３種類の試料孔を用い、ベ
ルオキシターゼ量を増加させて化学発光を行なわせ、各
試料孔での受光値を測定した。なお、各試料孔の側及び
底外面にはコーティング剤を塗布した。Experiment 2. Using three types of sample holes with light-harvesting bottom shapes of U-bottom, ■-bottom, and flat bottom, the amount of peroxidase was increased to cause chemiluminescence, and the light reception value at each sample hole was measured. Note that a coating agent was applied to the side and bottom outer surface of each sample hole.
標識酵素であるベルオキシターゼの量と発光量の関係は
直線的であることがすでに知られているが、本実験によ
れば、第６図に示すように、Ｕ底の試料孔ではＶ底、平
底の試料孔よりも集光性が良いため、ベルオキシターゼ
奄の変化に対する受光値の変化の勾配が大きく、感度が
良いことがわかる。It is already known that the relationship between the amount of peroxidase, which is a labeling enzyme, and the amount of luminescence is linear, but according to this experiment, as shown in Figure 6, in the U-bottom sample hole, the V-bottom, Since the light-gathering property is better than that of a flat-bottomed sample hole, the gradient of the change in the received light value with respect to the change in peroxidase volume is large, indicating that the sensitivity is good.
（発明の効果）
以上の説明から明らかなように、本発明に係る測定用プ
レートによれば、化学発光反応液が発する光の集光性が
良く、しかも外部光、特に隣接する試料孔での発光の影
響を受けないので、発光法による正確な測定が行なえる
。また、−度に多数の試料孔で同時に化学発光を行なわ
せることができ、迅速な測定が可能となるという効果を
有している。(Effects of the Invention) As is clear from the above description, the measurement plate according to the present invention has good condensing properties for the light emitted by the chemiluminescent reaction solution, and also has excellent light convergence properties, and is effective in reducing external light, especially in the adjacent sample holes. Since it is not affected by luminescence, accurate measurements can be made using the luminescence method. Furthermore, chemiluminescence can be caused to occur simultaneously in a large number of sample holes at the same time, which has the effect of enabling rapid measurement.
第１図は本発明に係る生物、化学発光酵素免疫測定用プ
レートの断面図、第２図は本発明の他の実施例に係る測
定用プレートの断面図、第３図は本発明に係る測定用プ
レートの斜視図、第４図は実験用の測定用プレートの平
面図、第５図は外部光の影響性に関する実験結果を示す
図、第６図は集光性に関する実験結果を示す図、第７図
は従来の測定用プレートの断面図である。
■・・・生物、化学発光酵素免疫測定用プレート、２・
・・試料孔、
３・・・コーティング剤。
特 許 出 願 人 帝國製薬株式会社代 理 人 弁
理士 青白 葆 ほか１名ｆｌｌｉ１ 図
第３図
寓２図
第７図
第４図FIG. 1 is a cross-sectional view of a plate for immunoassay of organisms and chemiluminescent enzymes according to the present invention, FIG. 2 is a cross-sectional view of a measurement plate according to another embodiment of the present invention, and FIG. FIG. 4 is a plan view of the experimental measurement plate, FIG. 5 is a diagram showing experimental results regarding the influence of external light, and FIG. 6 is a diagram showing experimental results regarding light gathering ability. FIG. 7 is a sectional view of a conventional measurement plate. ■...Biological, chemiluminescent enzyme immunoassay plate, 2.
...sample hole, 3...coating agent. Patent applicant: Representative of Teikoku Seiyaku Co., Ltd. Patent attorney: Aobai Ao and 1 other personflli1 Figure 3 Figure 2 Figure 7 Figure 4
Claims (4)
で形成するとともに、各試料孔の少なくとも側部及び底
部に非透光性処理を施したことを特徴とする生物、化学
発光酵素免疫測定用プレート。(1) An organism characterized by having a large number of sample holes with a bottom, the bottom of each sample hole being formed with a curved surface, and at least the sides and bottom of each sample hole being treated to make them non-transparent. Plate for chemiluminescent enzyme immunoassay.
又は黒色系塗料をコーティングしたことを特徴とする請
求項１に記載の生物、化学発光酵素免疫測定用プレート
。(2) The biological or chemiluminescent enzyme immunoassay plate according to claim 1, wherein at least the sides and bottom of the sample hole are coated with metal powder or black paint.
属粉末又は炭素粉末等を混入した合成樹脂で形成したこ
とを特徴とする請求項１に記載の生物、化学発光酵素免
疫測定用プレート。(3) The biological or chemiluminescent enzyme immunoassay plate according to claim 1, wherein at least the side and bottom portions of the sample hole are formed of a synthetic resin mixed with metal powder, carbon powder, or the like in advance.
成樹脂で形成したことを特徴とする請求項１に記載の生
物、化学発光酵素免疫測定用プレート。(4) The biological or chemiluminescent enzyme immunoassay plate according to claim 1, wherein at least the side and bottom portions of the sample hole are formed of a colored synthetic resin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1077529A JPH02254364A (en) | 1989-03-29 | 1989-03-29 | Plate for measuring immunity of biological/chemical luminescent enzyme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1077529A JPH02254364A (en) | 1989-03-29 | 1989-03-29 | Plate for measuring immunity of biological/chemical luminescent enzyme |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH02254364A true JPH02254364A (en) | 1990-10-15 |
Family
ID=13636509
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1077529A Pending JPH02254364A (en) | 1989-03-29 | 1989-03-29 | Plate for measuring immunity of biological/chemical luminescent enzyme |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH02254364A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06218304A (en) * | 1993-01-27 | 1994-08-09 | Asahi Sanac Kk | Paint spray machine for architecture |
| JP2011501132A (en) * | 2007-10-10 | 2011-01-06 | ポカード・ディアグノスティクス・リミテッド | System for identifying bacteria in urine |
| EP2931915A4 (en) * | 2012-12-11 | 2015-12-02 | Pocared Diagnostics Ltd | Optics cup with curved bottom |
| US9506866B2 (en) | 2008-02-05 | 2016-11-29 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in biological samples |
| US10288632B2 (en) | 2009-09-21 | 2019-05-14 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in biological samples |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59132335A (en) * | 1982-10-12 | 1984-07-30 | ダイナテク・ラボラトリ−ズ・インコ−ポレ−テツド | Non-fluorescent vessel for holding test sample for fluorometric analysis, its manufacture and fluorometric analysis method |
| JPS61215947A (en) * | 1985-03-22 | 1986-09-25 | Fujirebio Inc | Micro-plate for micro-titer |
| JPS6266141A (en) * | 1985-09-19 | 1987-03-25 | Sumitomo Bakelite Co Ltd | Vessel for fluorescent immunological measurement |
-
1989
- 1989-03-29 JP JP1077529A patent/JPH02254364A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59132335A (en) * | 1982-10-12 | 1984-07-30 | ダイナテク・ラボラトリ−ズ・インコ−ポレ−テツド | Non-fluorescent vessel for holding test sample for fluorometric analysis, its manufacture and fluorometric analysis method |
| JPS61215947A (en) * | 1985-03-22 | 1986-09-25 | Fujirebio Inc | Micro-plate for micro-titer |
| JPS6266141A (en) * | 1985-09-19 | 1987-03-25 | Sumitomo Bakelite Co Ltd | Vessel for fluorescent immunological measurement |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06218304A (en) * | 1993-01-27 | 1994-08-09 | Asahi Sanac Kk | Paint spray machine for architecture |
| JP2011501132A (en) * | 2007-10-10 | 2011-01-06 | ポカード・ディアグノスティクス・リミテッド | System for identifying bacteria in urine |
| US8808649B2 (en) | 2007-10-10 | 2014-08-19 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in urine |
| US9606105B2 (en) | 2007-10-10 | 2017-03-28 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in urine |
| EP3192876A1 (en) * | 2007-10-10 | 2017-07-19 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in urine |
| US10656140B2 (en) | 2007-10-10 | 2020-05-19 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in urine |
| US10801962B2 (en) | 2008-02-05 | 2020-10-13 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in biological samples |
| US9506866B2 (en) | 2008-02-05 | 2016-11-29 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in biological samples |
| US10073036B2 (en) | 2008-02-05 | 2018-09-11 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in biological samples |
| US10288632B2 (en) | 2009-09-21 | 2019-05-14 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in biological samples |
| US11002752B2 (en) | 2009-09-21 | 2021-05-11 | Pocared Diagnostics Ltd. | System for conducting the identification of bacteria in biological samples |
| US9862920B2 (en) | 2012-12-11 | 2018-01-09 | Pocared Diagnostics Ltd. | Optics cup with curved bottom |
| US10731123B2 (en) | 2012-12-11 | 2020-08-04 | Pocared Diagnostics Ltd. | Optics cup with curved bottom |
| EP2931915A4 (en) * | 2012-12-11 | 2015-12-02 | Pocared Diagnostics Ltd | Optics cup with curved bottom |
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