JPH01172755A - Method and apparatus for detecting abnormality of sealed immunoreaction reagent - Google Patents
Method and apparatus for detecting abnormality of sealed immunoreaction reagentInfo
- Publication number
- JPH01172755A JPH01172755A JP32996887A JP32996887A JPH01172755A JP H01172755 A JPH01172755 A JP H01172755A JP 32996887 A JP32996887 A JP 32996887A JP 32996887 A JP32996887 A JP 32996887A JP H01172755 A JPH01172755 A JP H01172755A
- Authority
- JP
- Japan
- Prior art keywords
- antigen
- antibody
- reaction
- fluorescence intensity
- test cup
- 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
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 29
- 230000005856 abnormality Effects 0.000 title claims abstract description 17
- 238000000034 method Methods 0.000 title claims abstract description 8
- 230000036046 immunoreaction Effects 0.000 title abstract description 21
- 238000012360 testing method Methods 0.000 claims abstract description 43
- 238000006243 chemical reaction Methods 0.000 claims abstract description 23
- 230000008105 immune reaction Effects 0.000 claims description 13
- 210000001124 body fluid Anatomy 0.000 claims description 7
- 239000010839 body fluid Substances 0.000 claims description 7
- 230000003287 optical effect Effects 0.000 claims description 3
- 230000002250 progressing effect Effects 0.000 claims description 2
- 238000005259 measurement Methods 0.000 abstract description 15
- 230000006866 deterioration Effects 0.000 abstract description 8
- 102000004190 Enzymes Human genes 0.000 abstract description 7
- 108090000790 Enzymes Proteins 0.000 abstract description 7
- 101100327917 Caenorhabditis elegans chup-1 gene Proteins 0.000 abstract description 6
- 239000000427 antigen Substances 0.000 abstract description 4
- 102000036639 antigens Human genes 0.000 abstract description 4
- 108091007433 antigens Proteins 0.000 abstract description 4
- 239000011324 bead Substances 0.000 abstract description 3
- 239000007788 liquid Substances 0.000 abstract description 3
- 238000012545 processing Methods 0.000 abstract description 2
- 238000001917 fluorescence detection Methods 0.000 description 7
- 239000000758 substrate Substances 0.000 description 6
- 102000003946 Prolactin Human genes 0.000 description 5
- 108010057464 Prolactin Proteins 0.000 description 5
- 229940097325 prolactin Drugs 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000004140 cleaning Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 238000006911 enzymatic reaction Methods 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000001174 ascending effect Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 238000009007 Diagnostic Kit Methods 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000005375 photometry Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
Abstract
Description
【発明の詳細な説明】
に適用される封入済み免疫反応試薬の異常を検出する方
法及び装置に関するものである。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method and apparatus for detecting abnormalities in encapsulated immune reaction reagents applied to.
免疫学的手法を利用した微量生体物質の測定、例えば診
断用キットとしての応用は近年高い注目を集め、様々な
試みがなされている。またそれにを得るためには装置の
安定性のみならず、使用する免疫反応試薬の品質が安定
していることが必要である。特に、予め免疫反応試薬を
封入したタイプのものは全数検査ができないため封入後
の免疫反応試薬の劣化等の異常については１００パーセ
ント保障することはできない。Measurement of trace amounts of biological substances using immunological techniques, such as application as diagnostic kits, has attracted a lot of attention in recent years, and various attempts have been made. In order to obtain this, it is necessary not only to have the stability of the device but also to have stable quality of the immunoreaction reagent used. In particular, since it is not possible to test all the products of the type in which the immunoreaction reagent is pre-encapsulated, it is not possible to guarantee 100% against abnormalities such as deterioration of the immunoreaction reagent after encapsulation.
本発明者がかかる観点から種々の検討を加えたところに
よると、抗原抗体反応進行中のテストカップ内に適当な
光源光を照射することにより、測定可能な蛍光強度が得
られ、更に劣化等の異常のある封入済み免疫反応試薬を
使用した場合、異常のない同一測定項目用の免疫反応試
薬を使用したである血清状び免疫反応試薬等に由来して
おり、その量に依存して増減するが、同一測定項目の場
合は使用される免疫反応試薬及び被検体液の量は一定で
あるた姥Ｍυ４冠となることを利用して免役反応試薬の
劣化等の異常を蛍光強度の変化として測定する方法を完
成させた。According to the inventor's various studies from this point of view, by irradiating an appropriate light source light into the test cup during the antigen-antibody reaction, measurable fluorescence intensity can be obtained, and furthermore, it is possible to prevent deterioration etc. If a prepackaged immune reaction reagent with an abnormality is used, it is derived from a serum or immune reaction reagent that was used for the same measurement item without an abnormality, and the amount will increase or decrease depending on the amount. However, in the case of the same measurement item, the amounts of immunoreaction reagent and sample body fluid used are constant, and by using this fact, abnormalities such as deterioration of the immunoreaction reagent can be measured as changes in fluorescence intensity. I have perfected the method.
本発明は抗原抗体反応の進行中にテストカップ内からの
蛍光強度を測定することにより、封入済み免疫反応試薬
の異常を検出する方法及び装置を提供するところにある
。The present invention provides a method and apparatus for detecting abnormalities in sealed immunoreaction reagents by measuring the fluorescence intensity from within the test cup during the progress of antigen-antibody reaction.
前記の目的を達成するためになされた本発明の特徴は、
予めテストカップに封入された免疫反応試薬に被検体液
を添加後、テストカップの中で抗原抗体反応の進行中に
テストカップ内からの蛍光強度を測定し、標準値と比較
するところにある。The features of the present invention achieved to achieve the above object are as follows:
After adding a sample body fluid to an immunoreaction reagent previously sealed in a test cup, the fluorescence intensity from within the test cup is measured while the antigen-antibody reaction is progressing in the test cup, and compared with a standard value.
また、かかる方法を好適に実現するための本発明よりな
る装置の特徴は、免疫反応測定装置において、抗原抗体
反応の反応室な提供するテストカップ内からの蛍光強度
を測定する光学手段を抗原抗体反応の進行中の蛍光強度
が測定可能な位置に設置した所にある。Further, the feature of the apparatus according to the present invention for suitably realizing such a method is that in an immunoreaction measuring apparatus, an optical means for measuring the fluorescence intensity from within a test cup provided in a reaction chamber for antigen-antibody reaction is used as a reaction chamber for antigen-antibody reaction. It is located at a location where the fluorescence intensity during the reaction can be measured.
体反応進行中のテストカップ内から得られる蛍光強度は
同一測定項目の場合はほぼ一定となるが、使用する免疫
反応試薬に劣化等の異常がある場合−（試料？
した場合正しい測定結果を不した弐鋸がその抗原さらに
、被検体液添加後の抗原抗体反応進行中に測定するのは
、例えば免疫反応試薬が予め凍結乾燥等されてテストカ
ップに填加されている場合に、免疫反応試薬が溶解し、
蛍光を発する様な状態にして測定するためである。The fluorescence intensity obtained from inside the test cup during a body reaction is almost constant for the same measurement item, but if there is an abnormality such as deterioration in the immunoreaction reagent used, the correct measurement result may be incorrect. The antigen is then measured during the antigen-antibody reaction after addition of the sample body fluid. For example, if the immune reaction reagent has been freeze-dried in advance and loaded into the test cup, the immunoreaction reagent is dissolves,
This is to perform measurements in a state where fluorescence is emitted.
本発明において、蛍光強度を測定するための励起波長及
び測定波長は測定に充分な蛍光強度の変化が得られれば
どのような波長でも制限はな（、例えば、励起波長５６
０ ｎｍ測定波長４５０ ｎｎ＋で測定できる。In the present invention, the excitation wavelength and measurement wavelength for measuring fluorescence intensity are not limited to any wavelength as long as a sufficient change in fluorescence intensity is obtained for measurement (for example, excitation wavelength 56
Can be measured at 0 nm measurement wavelength 450 nn+.
本発明装置において、用いられる光学手段は、公知の蛍
光検出装置等で良く、またそれらはテストカップ内から
の蛍光を測定可能な場所に設置されていれば良い。In the apparatus of the present invention, the optical means used may be a known fluorescence detection device or the like, and it is sufficient that they are installed at a location where fluorescence from within the test cup can be measured.
本発明は、封入済み免疫反応試薬を使用する免疫反応測
定装置に適用され、例えば具体的に例示すれば、特開昭
６′２−１４８８５８号に記載の装置等に適用される。INDUSTRIAL APPLICABILITY The present invention is applied to an immunoreaction measuring device using an encapsulated immunoreaction reagent, for example, to the device described in Japanese Patent Application Laid-Open No. 6'2-148858.
以下本発明の一実施例について図面にもとすき説明する
。An embodiment of the present invention will be described below with reference to the drawings.
第１図に示された公知の蛍光検出装置は、被検体液と免
疫反応試薬等の反応室であるテストカップ１に対し、光
源２からの光源光を、励起側フィルタ３ヶ通しダイクロ
イックミラー４から集光レラー４、受光側フィルタ６を
経て受光センサ７で受光し、所定の信号処理回路（図示
せず）で蛍光強度を測定するようになっている。In the known fluorescence detection device shown in FIG. 1, light source light from a light source 2 is passed through three excitation-side filters to a test cup 1, which is a reaction chamber containing a sample body fluid, an immune reaction reagent, etc., and a dichroic mirror 4. The light is received by a light receiving sensor 7 through a condensing mirror 4 and a light receiving filter 6, and the fluorescence intensity is measured by a predetermined signal processing circuit (not shown).
第２図は自動酵素免疫反応測定装置に適用した場合の構
成概９を示した図であり、９は所定のテストカップをテ
ストプレートに組込み搭載するテストカップストレージ
部であり、エレベータ機構１０．１）の間に搬送経路１
２が設けられ、この搬送経路の上流から下流に向かって
、テストカップの上方間シールを挿装させるシールブレ
イク装［１５、被検体液分注装置１４、試薬の異常検出
用の蛍光検出装置１５、Ｂ／Ｆ分離および洗浄装置１６
、基質溶液注入装置１７、酵素反応に由来して得られた
試料中の蛍光物質の蛍光強度測定用の蛍光検出装置１８
が、所定の間隔で搬送経路の上側に設けられている。FIG. 2 is a diagram showing an outline of the configuration 9 when applied to an automatic enzyme immunoreaction measuring device, in which 9 is a test cup storage section in which a predetermined test cup is assembled and mounted on a test plate, and an elevator mechanism 10.1 ) between transport path 1
2, a seal breaking device [15] for inserting the upper seal of the test cup from upstream to downstream of the transport path, a sample liquid dispensing device 14, and a fluorescence detection device 15 for detecting abnormalities in the reagent. , B/F separation and cleaning device 16
, a substrate solution injection device 17, and a fluorescence detection device 18 for measuring the fluorescence intensity of a fluorescent substance in a sample obtained from an enzyme reaction.
are provided above the conveyance path at predetermined intervals.
以上のような構成の自動酵素免疫反応測定装置において
は、まずテストカップストレージ部９で所定の数、組合
わせのテストカップを組込み搭載したテストプレートＹ
、上昇用エレベータ１０かも搬送経路１２の上流位置に
押しあげる。次いで押し込みシリンジ装置により搬送経
路１２にテストプレートを押し込みすると、以後は、ラ
チェツト爪をもつ間欠搬送手段により、所定のタイミン
グ間隔で一定長ずつテストプレートは下流側に送られる
ことになる。In the automatic enzyme immunoreaction measuring device having the above configuration, first, the test plate Y on which a predetermined number and combination of test cups are installed is stored in the test cup storage section 9.
, the ascending elevator 10 is also pushed up to the upstream position of the conveyance path 12. Next, when the test plate is pushed into the conveyance path 12 by the pushing syringe device, the test plate is thereafter conveyed downstream by a constant length at predetermined timing intervals by an intermittent conveyance means having a ratchet pawl.
このテストプレートは、まずシールブレイク装置１３の
位置に移入されて、上下動式くさびの下動によりテスト
カップ上面のシール乞押し込みブレイクする。シールブ
レイクされたテストカップは、試料分注装置１４の位置
に移入され、所定の被検体液が各テストカップ内に添加
される。本例のテストカップ１は、表面に抗体の結合さ
れている磁性ビーズ８と、酵素の標識された別の抗体と
が予め内部に填加されているものであり、前記抗原を含
む被検体液の添加により固相化抗体−抗原−酵素標識抗
体の特異的反応が行われる。この反応は一定時間行われ
ることが必要であり、従って本例では第２図に示す如く
搬送経路１２に所定長のインキュベート領域を与えてい
る。This test plate is first moved to the position of the seal breaking device 13, and the seal on the top surface of the test cup is forced and broken by the downward movement of the vertically movable wedge. The test cups whose seals have been broken are transferred to the position of the sample dispensing device 14, and a predetermined body fluid to be tested is added into each test cup. The test cup 1 of this example has magnetic beads 8 to which an antibody is bound on the surface and another enzyme-labeled antibody loaded in advance, and the test cup 1 contains a sample liquid containing the antigen. By adding , a specific reaction between the immobilized antibody-antigen-enzyme-labeled antibody is carried out. This reaction needs to be carried out for a certain period of time, and therefore, in this example, an incubation region of a certain length is provided in the transport path 12 as shown in FIG.
テストプレートがこのインキュベート領域を通−兜豊長
±比較することにより、
免疫反応試薬の異常の有無を確めることができる。By comparing the length of the test plate passed through this incubation area, it is possible to confirm whether or not there is an abnormality in the immunoreaction reagent.
抗原抗体反応が充分になされたテストカップを搭載した
テストプレートは、Ｂ／Ｆ分離および洗浄装置１６の位
置に移入され、Ｂ／Ｆ分離、洗浄がなされる。The test plate loaded with the test cup in which the antigen-antibody reaction has been sufficiently performed is transferred to the B/F separation and cleaning device 16, where it is subjected to B/F separation and cleaning.
次いでテストプレートは基質溶液注入装置１７に移入さ
れ、テストカップに基質溶液が添加され、酵素反応が開
始される。The test plate is then transferred to the substrate solution injection device 17, the substrate solution is added to the test cup, and the enzyme reaction is started.
基質溶液の添加されたテストカップは、蛍光検出装置１
８の位置に移入され、基質に生じた変化状態が光学的に
測定され、この測定情報にもとすき被検体液中の抗原濃
度が定量される。The test cup to which the substrate solution has been added is placed in the fluorescence detection device 1.
8, the state of change that occurs in the substrate is optically measured, and the antigen concentration in the sample body fluid is quantified based on this measurement information.
測光が終了したテストプレートは下降用エレベータによ
りテストプレートストレージ部９に移送される。The test plate on which photometry has been completed is transferred to the test plate storage section 9 by a descending elevator.
このような構成によれば、測定を続けながらその免疫反
応試薬の劣化等の異常を検出することが可能であり、測
定結果の信頼性が向上するという効果がある。According to such a configuration, it is possible to detect abnormalities such as deterioration of the immune reaction reagent while continuing the measurement, and the reliability of the measurement results is improved.
実施例に示した装置及びそれ専用の封入済み酵素免疫反
応試薬で使用期限を過ぎたため劣化した可能性のあるも
のを用いて、２種類の血清中のプロラクチン（ＰＲＬ）
’＆連続して複数回測定した場合のＰＲＬの定量結果と
抗原抗体反応進行中の蛍光強度の関係を第１表に示した
。酵素免疫反応試薬の劣化のためＰＲＬの定量結果が期
待値から大ぎく外れている場合の抗原抗体反応進行中の
蛍光強度は、ＰＲＬの定量結果が期待値に近い場合に比
べて、有為に増加している。Prolactin (PRL) in two types of serum was measured using the device shown in the example and its dedicated enzyme immunoreaction reagent, which may have deteriorated because it had passed its expiration date.
Table 1 shows the relationship between the quantitative results of PRL and the fluorescence intensity during the progress of the antigen-antibody reaction when measured multiple times in succession. When the PRL quantification result deviates significantly from the expected value due to deterioration of the enzyme immunoreaction reagent, the fluorescence intensity during the antigen-antibody reaction will be significantly lower than when the PRL quantification result is close to the expected value. It has increased.
本発明より、抗原抗体反応進行中の蛍光強度を同−測定
項目間で比較することにより、免疫反応測定を行いなが
ら封入済み免疫反応試薬の異常を検出することが可能に
なった。また、劣化等の異常のある免疫反応試薬による
定量結果を除くことにより、より信頼性が筒い定ｉ？：
行うことができるようになった。According to the present invention, by comparing the fluorescence intensity during the progress of antigen-antibody reaction between the same measurement items, it has become possible to detect abnormalities in the encapsulated immune reaction reagent while performing immune reaction measurement. In addition, by excluding quantitative results from immunoreaction reagents with abnormalities such as deterioration, reliability can be increased. :
Now you can do it.
第１図は本発明の一実施例を説明するための蛍光測定装
置の構成概要を示した図、第２図は自動酵素免疫測定装
置に適用した場合の構成概要を示した図である。
１・・・テストカップ
２・・・光諒
３・・・励起側フィルタ
４・・・ダイクロイックミラ−
５・・・集光レンズ
６・・・受光側フィルタ
７・・・受光センサ
８・・・磁性ビーズ
９・・・テストカップストレージ
１０・・・上昇用エレベータ
１）・・・下降用エレベータ
１２・・・搬送経路
１３・・・シールブレイク装置
１４・・・被検体液分注装置
１５・・・異常免疫反応試薬検出用蛍光検出装置１６・
・・Ｂ／Ｆ分離及び洗浄装置
１７・・・基質溶液注入装置
１８・・・酵素反応に由来して得られた蛍光物質測定用
蛍光検出装置
ｌ−７入ト刀ツノFIG. 1 is a diagram showing an outline of the configuration of a fluorescence measuring device for explaining an embodiment of the present invention, and FIG. 2 is a diagram showing an outline of the configuration when applied to an automatic enzyme immunoassay device. 1... Test cup 2... Light beam 3... Excitation side filter 4... Dichroic mirror 5... Condensing lens 6... Light receiving side filter 7... Light receiving sensor 8... Magnetic beads 9... Test cup storage 10... Ascending elevator 1)... Descending elevator 12... Transport path 13... Seal breaking device 14... Subject fluid dispensing device 15...・Fluorescence detection device 16 for detecting abnormal immune reaction reagents・
...B/F separation and cleaning device 17...Substrate solution injection device 18...Fluorescence detection device l-7 for measuring fluorescent substances derived from enzyme reactions
Claims (2)
検体液を添加後、テストカップの中で抗原抗体反応の進
行中にテストカップ内からの蛍光強度を測定し、標準値
と比較することを特徴とするの封入済み免疫反応試薬の
異常を検出する方法。(1) After adding the test body fluid to the immune reaction reagent sealed in the test cup in advance, measure the fluorescence intensity from inside the test cup while the antigen-antibody reaction is progressing in the test cup, and compare it with the standard value. A method for detecting an abnormality in an encapsulated immune reaction reagent, characterized by:
室を提供するテストカップ内からの蛍光強度を測定する
光学手段を抗原抗体反応の進行中の蛍光強度が測定可能
な位置に設置したことを特徴とする封入済み免疫反応試 薬の異常を検出する装置。(2) In the immune reaction measuring device, the optical means for measuring the fluorescence intensity from inside the test cup, which provides the reaction chamber for the antigen-antibody reaction, is installed at a position where the fluorescence intensity during the progress of the antigen-antibody reaction can be measured. A device that detects abnormalities in sealed immune reaction reagents.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP32996887A JPH01172755A (en) | 1987-12-28 | 1987-12-28 | Method and apparatus for detecting abnormality of sealed immunoreaction reagent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP32996887A JPH01172755A (en) | 1987-12-28 | 1987-12-28 | Method and apparatus for detecting abnormality of sealed immunoreaction reagent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01172755A true JPH01172755A (en) | 1989-07-07 |
Family
ID=18227281
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP32996887A Pending JPH01172755A (en) | 1987-12-28 | 1987-12-28 | Method and apparatus for detecting abnormality of sealed immunoreaction reagent |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01172755A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011064778A3 (en) * | 2009-11-30 | 2011-08-04 | Bio-Rad Laboratories Inc. | Optical bead assay reader |
-
1987
- 1987-12-28 JP JP32996887A patent/JPH01172755A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011064778A3 (en) * | 2009-11-30 | 2011-08-04 | Bio-Rad Laboratories Inc. | Optical bead assay reader |
| US8927294B2 (en) | 2009-11-30 | 2015-01-06 | Bio-Rad Laboratories Inc. | Bead reader |
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