JPH0287063A - Determining method of prozone in immunoreaction - Google Patents
Determining method of prozone in immunoreactionInfo
- Publication number
- JPH0287063A JPH0287063A JP23924888A JP23924888A JPH0287063A JP H0287063 A JPH0287063 A JP H0287063A JP 23924888 A JP23924888 A JP 23924888A JP 23924888 A JP23924888 A JP 23924888A JP H0287063 A JPH0287063 A JP H0287063A
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- JP
- Japan
- Prior art keywords
- sample
- antigen
- antibody
- reaction
- reagent
- 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.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 33
- 230000036046 immunoreaction Effects 0.000 title 1
- 238000006243 chemical reaction Methods 0.000 claims abstract description 47
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 38
- 238000002835 absorbance Methods 0.000 claims abstract description 28
- 239000000427 antigen Substances 0.000 claims abstract description 18
- 102000036639 antigens Human genes 0.000 claims abstract description 18
- 108091007433 antigens Proteins 0.000 claims abstract description 18
- 239000002245 particle Substances 0.000 claims description 11
- 239000012085 test solution Substances 0.000 claims description 5
- 229940127121 immunoconjugate Drugs 0.000 claims description 3
- 239000000243 solution Substances 0.000 claims description 3
- 230000001678 irradiating effect Effects 0.000 claims description 2
- 230000008105 immune reaction Effects 0.000 claims 2
- 238000005259 measurement Methods 0.000 abstract description 9
- 230000008569 process Effects 0.000 abstract description 5
- 230000007246 mechanism Effects 0.000 description 10
- 239000000126 substance Substances 0.000 description 9
- 210000002966 serum Anatomy 0.000 description 7
- 238000004140 cleaning Methods 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 229940079593 drug Drugs 0.000 description 5
- 238000003860 storage Methods 0.000 description 4
- 230000004520 agglutination Effects 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 239000011810 insulating material Substances 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005375 photometry Methods 0.000 description 1
- 238000013102 re-test Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000000439 tumor marker Substances 0.000 description 1
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
Abstract
Description
【発明の詳細な説明】
（イフ産業上の利用分野
この発明は抗原抗体反応が抗原過剰域および抗体過剰域
のいずれで行なわれているか全判定（１０ゾーン判定〕
する方法に関するＯ
Ｐ）従来の技術
体液中の被測定物質（抗原または抗体）の測定法として
被測定物質と抗原抗体反応しうる物質（抗体または抗原
）を作用させ、生じる抗原抗体結合物による凝集の度合
全測定する方法がある。Detailed Description of the Invention (IF Industrial Application Field) This invention is a complete determination of whether the antigen-antibody reaction is occurring in the antigen-excessive region or the antibody-excessive region (10-zone determination)
O P) Conventional technology A method for measuring a substance to be measured (antigen or antibody) in a body fluid is to react with a substance (antibody or antigen) capable of antigen-antibody reaction with the substance to be measured, and agglutination is caused by the resulting antigen-antibody combination. There is a way to fully measure the degree of
これには１例えば、上記被測定物質を含む検体と上記抗
原抗体反応しうる物質とを直接反応させる免疫比濁法や
、上記抗原抗体反応しうる物質を不溶性坦体に坦持させ
た試薬全検体に作用さぜる方決（ｆｌｌえはラテックス
凝集反応性）がある。いずれの場合にも、生じた凝集の
度合を、被検液に元を照射して、透過光強度の減衰や散
乱光強度の増加を測定する。Examples of this include immunoturbidimetry in which a sample containing the above-mentioned substance to be measured is directly reacted with the above-mentioned substance capable of reacting with the antigen-antibody, and whole reagents in which the above-mentioned substance capable of reacting with the antigen-antibody is supported on an insoluble carrier. There is a method of acting on the specimen (the latter is latex agglutination reactivity). In either case, the degree of aggregation that has occurred is determined by irradiating the test liquid with a source and measuring the attenuation of the transmitted light intensity and the increase in the scattered light intensity.
例えば第４図に示すように、血清中に含１れる免疫グロ
ブリンＩｇＧｌｊ、血清と抗１ｇＧ抗体を含む溶液とを
混合し、生じた抗原抗体結合物の量を適当な波長におけ
る吸光度全測足することによＶ測定される。この抗原抗
体結合物生成量はＩｇＧ濃度が高くなるにつれて多くな
り、かつ結合物の粒子径が大きくなるため吸光度が上昇
する。しかしながらＩｇＧ濃度がある値以上になると抗
原による抗原抗体架橋効果がなくなるため、結合物の粒
子径が小さくなり、その結果吸光度が低下する。このた
め、ある吸光度に対するＩｇＧ ｆｉ度が一鵜的に定ｌ
らない、これをプロゾーン現象とよび、抗原抗体反応が
抗原過剰域および抗体過剰域のいずれで行なわれている
か全判定（プロゾーン判定）する必要がある。For example, as shown in Figure 4, immunoglobulin IgGlj contained in serum, serum and a solution containing anti-1gG antibody are mixed, and the amount of antigen-antibody conjugate produced is measured by total absorbance at an appropriate wavelength. In particular, V is measured. The amount of the antigen-antibody conjugate produced increases as the IgG concentration increases, and the absorbance increases as the particle size of the conjugate increases. However, when the IgG concentration exceeds a certain value, the antigen-antibody crosslinking effect by the antigen disappears, so the particle size of the conjugate decreases, resulting in a decrease in absorbance. For this reason, the IgG fi degree for a certain absorbance is almost constant.
This is called the prozone phenomenon, and it is necessary to make a complete determination (prozone determination) as to whether the antigen-antibody reaction is occurring in the antigen-excessive region or the antibody-excessive region.
プロゾーン判定の方法として例えば、特開昭６０７９２
６９号にμ検体の量を変えて判定する方法が開示されて
いる。このように異った濃度で複数回反応させることに
より、最初の反応が抗原過剰域か抗体過剰域かが判るが
、検体の希釈など工程が複雑になると同時に、高価な試
薬が倍必要であるという経済上の問題もある。このため
１反応が終了した被検液に、測定目的成分（抗原または
抗体）を含む標準液を添加して、凝集が更に進む（吸光
度が上昇する）かどうかを見て、抗原過剰域か抗体過剰
域かを判定する方法も提案されているが、この場合には
濃度の測定結果を出した後、被検液を回収したり９反応
管を直接測光したすすることが必要であり、かつ判定の
ための反応時間が必要であるといり問題点があり、測定
を自動化する場合、装置が複雑になる。For example, as a method of pro zone determination, JP-A No. 60792
No. 69 discloses a method for determination by changing the amount of μ specimen. By reacting multiple times at different concentrations in this way, it can be determined whether the initial reaction is in the antigen-rich region or in the antibody-rich region, but this complicates the process, such as diluting the sample, and requires twice as many expensive reagents. There is also an economic problem. For this reason, add a standard solution containing the component to be measured (antigen or antibody) to the test solution after one reaction, and check whether the agglutination progresses further (absorbance increases). A method for determining whether the concentration is in the excess range has also been proposed, but in this case, it is necessary to collect the test solution or directly photometer the reaction tube and rinse it after obtaining the concentration measurement result. There are problems in that a reaction time is required for the determination, and if the measurement is automated, the equipment becomes complicated.
筐た９例えば特公昭６１−１０７７５号に示されている
ように、抗原抗体反応全経時的に追跡し反応進行のパタ
ーン力・ら抗原過剰域か抗体過剰域かを判定する方法や
、特開昭６３−１９５６０号に示されているように、二
つの波長での吸光度の比をとることにより１粒子径の大
きさ全判定し、抗原過剰域か抗体過剰域か全判定する方
決が示されている。For example, as shown in Japanese Patent Publication No. 61-10775, there is a method of tracing the antigen-antibody reaction over time and determining whether it is in an antigen-excessive region or an antibody-excessive region based on the pattern of the reaction progress, and As shown in No. 63-19560, a method is shown in which the size of a single particle can be completely determined by taking the ratio of the absorbance at two wavelengths, and whether it is an antigen-excessive area or an antibody-excessive area. has been done.
Ｖつ発明が解決しようとする問題点
しかしながら、ｇ＆光度の経時変化をみる方法や１粒子
径を判定する方法では、おる程度の抗原過剰までは判定
できるが抗原が大過剰の場合には。V Problems to be Solved by the Invention However, with the method of looking at changes in g&luminosity over time and the method of determining the diameter of a single particle, it is possible to determine if there is a slight excess of antigen, but when there is a large excess of antigen.
抗原抗体反応による結合物の凝集が起らないために、抗
原が非常に低濃度の場合との差を判定することがむつか
しいという問題点がある。例えば。There is a problem in that it is difficult to judge the difference from a case where the antigen is at a very low concentration because aggregation of the bound product does not occur due to the antigen-antibody reaction. for example.
腫瘍マーカと呼ばれるα−フ二トプロテイン（ＡＦＰ）
は、正常な人では数１０ ｎｇ／ｍ！であるが。α-phnitoprotein (AFP), called a tumor marker
is several tens of ng/m in a normal person! In Although.
原発性肝癌の、＠＠では数１００．０００ｎｇ／ｒｎ／
になること％あるといわれている。For primary liver cancer, @@ is several 100.000ng/rn/
It is said that there is a possibility of becoming
この発明は、かかる状況に鑑みなされたものであり、こ
とに抗原大過剰の場合にもプロゾーン判定を容易、かつ
確寮で経済的に行なうことの可能なプロゾーン判定方法
を提供しようとするものである。This invention has been made in view of the above situation, and aims to provide a prozone determination method that allows prozone determination to be performed easily and economically even in the case of a large excess of antigens. It is something.
に）問題点全解決するための手段
この発明の抗原抗体反応のプロゾーン判％［μ 審
ｍＪ試料（抗原または抗体を含む体液）と該目的成分を
含む標準試料の両方全反応容器に分注する工程。2) Means for solving all the problems Prozone test percentage of antigen-antibody reaction of this invention [μ] Dispense both the sample (body fluid containing antigen or antibody) and the standard sample containing the target component into all reaction vessels. The process of doing.
（ｂ）上記に試薬（抗体または抗原）ｋ添加して。(b) Add reagent (antibody or antigen) to the above.
抗原抗体反応を行なわせる工程。The process of causing an antigen-antibody reaction.
ｔｃノ反応の終末あるいは過程における透過光あるいは
散乱光強度を測定する工程。A step of measuring the intensity of transmitted light or scattered light at the end or process of the tc reaction.
（ｄ）上記の測定値をｔとに、試料中の目的成分濃度を
算出すると同時に、抗原過剰域か抗体過剰域かを判定す
る工程。(d) Using the above measured value as t, calculating the concentration of the target component in the sample and simultaneously determining whether it is in the antigen-excessive region or the antibody-excessive region.
から成立つ。It is established from
この発明の方法の最も特徴とする点は、に料に標準試料
を添加し、目的成分の濃度を上げて反応を行なわせ、標
準試料のみに試薬を加えて測定した際の吸光度あるいは
散乱光強度の値以下では目的成分のｌｌｌｋ度が大過剰
であると判定し、その値以上では反応進行のパターンの
データあるいは二つの波長の吸光度比のデータにより、
抗原過剰域か抗体過剰域かの判定を行なう点である◇例
えば、第２図はＩｇＧ濃度既知の血清（約７０００ｍｇ
／ｄｅ）を希釈したものの一定量に標準試料（コントロ
ール血清）の一定量を添加したものと試薬（ＩｇＧ抗血
清）全反応させ、血清中のＩｇＧ濃度と吸光度の関係を
示したものである。The most distinctive feature of the method of this invention is that a standard sample is added to the food, the concentration of the target component is increased, the reaction is carried out, and the absorbance or scattered light intensity is measured by adding a reagent only to the standard sample. Below this value, it is determined that the target component has a large degree of excess, and above that value, based on data on the pattern of reaction progress or data on the absorbance ratio of two wavelengths,
This is the point to judge whether it is an antigen-excessive area or an antibody-excessive area.
The figure shows the relationship between the IgG concentration in the serum and the absorbance when a reagent (IgG antiserum) was reacted with a certain amount of a standard sample (control serum) added to a certain amount of a diluted version of ``/de''.
濃度未知の検体について、上記と同様の手順で反応させ
た時の吸光度が第２図の点線以下であれば、 ＩｇＧが
大過剰（６００Ｑ ｍｇ；／ｄ ｅ以上）と判断し。When a sample with an unknown concentration is reacted using the same procedure as above, if the absorbance is below the dotted line in Figure 2, it is determined that IgG is in large excess (more than 600 Q mg;/de).
例えば検体２１０倍希釈して再＠する。また吸光度が第
２図の点線以上であれば１例えば第３肉に示すよ’）
ＩＣ３４０ｎｍの吸光度Ａ３４０と７００ｎｍＬｖ［ｉ
光度Ａ７００（７）比Ａ３４０／Ａ７００ ＶＣよって
抗体過剰域か抗原過剰域か全判定する。第３図に示すよ
うな吸光度比だけで抗体過剰域か抗原過剰域か全判定し
ようとした場合、超高濃度域（６０００ｍｇ／ｄｅ以上
〕の判定ができないが１本発明では、標準試料の添加に
より、予め超高濃度かどうかを判定しているので９両者
の組合わせにより全濃度域全カバーできることが特徴で
ある。For example, dilute the sample 210 times and re-test. Also, if the absorbance is above the dotted line in Figure 2, it will be shown as 1, for example, in the 3rd line.')
Absorbance A340 of IC340nm and 700nmLv[i
Based on the luminous intensity A700 (7) ratio A340/A700 VC, it is determined whether it is in the antibody excess region or the antigen excess region. If one attempts to determine whether the antibody-excessive region or antigen-excessive region is present only by the absorbance ratio as shown in Figure 3, it is not possible to determine the ultra-high concentration region (6000 mg/de or more). Since it is determined in advance whether or not the concentration is ultra-high, the combination of both nine can cover the entire concentration range.
標準試料中の目的成分の量が少な過ぎると１両者の組合
わせによっても判定で′ｆ！ない領域が生ずるし、目的
成分の量が多すぎると、早くプロシンを生じることＫな
９．測定範囲が狭くなるので。If the amount of the target component in the standard sample is too small, it will be judged by the combination of the two. 9. If the amount of the target component is too large, pro-syn will be generated quickly.9. Because the measurement range becomes narrower.
目的成分および使用する試薬に応じて、添加量全選定す
ることが重要である。It is important to select the total amount to be added depending on the target component and the reagent used.
この発明の方法は１通常の分光光度計を用いて用手法で
行なうこともできるが９反応管直接測光方式で反応過程
の吸光度データ全測定することができ、さらに２つの波
長の吸光度の死金出力できる自動化学分析装置を用いて
行なうのが適している。Although the method of this invention can be carried out manually using a normal spectrophotometer, it is also possible to measure all the absorbance data of the reaction process using the reaction tube direct photometry method. It is suitable to perform this using an automatic chemical analyzer that can output.
（ホ）作用ならびに実施例
この発明によれば、ＦＳ科に該目的成分を含む標準試料
を添加した後、ＶＣ薬と抗原抗体反応を行なわせるので
、試料中に目的成分が少ししか含まれていない場合でも
適量の抗原抗体結合粒子を生成させることができる。(E) Effects and Examples According to the present invention, after adding a standard sample containing the target component to the FS sample, an antigen-antibody reaction with the VC drug is performed, so that the sample contains only a small amount of the target component. Even in the absence of such particles, a suitable amount of antigen-antibody binding particles can be produced.
また、試料中に目的成分が大過剰に含まれているため適
量の抗原抗体結成粒子上生成することができない場合を
区別して検知することができる。In addition, it is possible to distinguish and detect cases where the target component is contained in a large excess in the sample and cannot be produced on the antigen-antibody forming particles in an appropriate amount.
それ以外の適量の抗原抗体結合粒子を生成した場合につ
いては１粒子生成による吸光度の経時的変化のパターン
や９反応後の生成粒子の平均的な大きさ（２つの波長の
吸光度比）を測定することにより、抗体過剰域での反応
であるか、抗原過剰域での反応であるか全区別すること
ができる。 Ｌ７Ｃがって添加する標準試料の濃度ある
いけ添加量全選定することにより、試料中の目的成分の
あらゆる濃度域について、容易かつ確突で経済的なプロ
ゾーン判定を行なうことが可能となる。In other cases where an appropriate amount of antigen-antibody binding particles are generated, measure the pattern of change in absorbance over time due to the generation of one particle and the average size of the particles generated after 9 reactions (absorbance ratio of two wavelengths). By doing so, it is possible to completely distinguish whether the reaction is in the antibody-excessive range or the antigen-excessive range. By selecting the concentration and amount of the standard sample to be added according to L7C, it becomes possible to easily, accurately, and economically perform prozone determination for all concentration ranges of the target component in the sample.
以下、第１図を参照しながら本発明方決全説明する。Hereinafter, the method of the present invention will be fully explained with reference to FIG.
第１図は、この発明の方法の突施に用いる測定装置の一
例の構成説明図である。第１図において１は試料分注ポ
ンプ、２は試料分注ノズル、３は試料分注ノズル移動機
摺、４・５はそれぞれ標準試料容器および標準試料、６
は試料用ターンテープ／Ｌ’、７・８セそれぞれ試料容
器および試料、９は反応ディスク、 １０． （１０’
・１０′）は反応セル。FIG. 1 is an explanatory diagram of the configuration of an example of a measuring device used in the method of the present invention. In Fig. 1, 1 is a sample dispensing pump, 2 is a sample dispensing nozzle, 3 is a sample dispensing nozzle moving mechanism slide, 4 and 5 are a standard sample container and a standard sample, respectively, and 6
10. Turn tape for sample/L', 7th and 8th cell respectively sample container and sample, 9 reaction disk, 10. (10'
・10') is a reaction cell.
１１は第１ｖＣ薬分注ポンプ、１２は第１試薬分注ノズ
ル、１３は第１ｖＳ薬分注ノズル移動機構、１４は試薬
（≦）ノ
庫、１５・１６はそれぞれ第１試薬容器および第１試薬
、１７は分光器、１８は分光器移動機構、１９は制御お
よびデータ処理コンピュータ、２０は第２に薬分性ポン
プ、２１は第２試薬分注ノズル、２２は第２試薬分注ノ
ズｐ移動機構、２３・２４ハそれぞれ第２試薬容器およ
び第２に薬、２５ハ洗浄ポンプ、２６は洗浄ノズル上下
機構、２７は洗浄ノズルでおる。11 is a first vC drug dispensing pump, 12 is a first reagent dispensing nozzle, 13 is a first vS drug dispensing nozzle movement mechanism, 14 is a reagent (≦) storage, and 15 and 16 are a first reagent container and a first reagent dispensing nozzle, respectively. Reagent, 17 is a spectrometer, 18 is a spectrometer moving mechanism, 19 is a control and data processing computer, 20 is a second chemical dispensing pump, 21 is a second reagent dispensing nozzle, 22 is a second reagent dispensing nozzle p A moving mechanism, 23 and 24 respectively a second reagent container and a second medicine, 25 a cleaning pump, 26 a cleaning nozzle up/down mechanism, and 27 a cleaning nozzle.
かかる装置において、試料分注ポンプ１と連結されてい
る試料分注ノズ／Ｉ／２が試料分注ノズル移動機構３に
よって移動し、標準試料容器４から一定量の標準試料５
會吸引し、続いて試料用ターンテープｌｖ６に七ツ）さ
れた試料容器７から一定量の試料８を吸引し１反応ディ
スク９に配置されている反応セル１０の中に試料８およ
び標準試料５を分注する。反応デイヌク９が回転して反
応上Ｉｖ１０が１ステップ進んだところで、第１試薬分
注ポンプ１１と連結されている第１試薬分注ノズＩｖ１
２が第１Ｋ薬分注ノズル移動機構１３によって移動し、
試薬庫１４内にホットされている第１試薬容器１５から
一定量の第１試薬１６を吸引し、続いて反応十ル１０α
α
のところに移動して反応セ／Ｌ／１σ内に分注する。In this device, a sample dispensing nozzle /I/2 connected to a sample dispensing pump 1 is moved by a sample dispensing nozzle moving mechanism 3, and a fixed amount of a standard sample 5 is transferred from a standard sample container 4.
Then, a certain amount of sample 8 is aspirated from the sample container 7 which is attached to the sample turn tape lv6, and the sample 8 and standard sample 5 are placed in the reaction cell 10 arranged on one reaction disk 9. Dispense. When the reaction valve 9 rotates and the reaction Iv10 advances by one step, the first reagent dispensing nozzle Iv1 connected to the first reagent dispensing pump 11
2 is moved by the first K drug dispensing nozzle moving mechanism 13,
A certain amount of the first reagent 16 is sucked from the first reagent container 15 heated in the reagent storage 14, and then the reaction container 10α
Move to α and dispense into reaction cell/L/1σ.
分光器１７が分光器移動機構１８により反応ディスク９
と同じ軸の回りに往復回転しながら各反応上μについて
、２つの波長λ１・λ２での吸光度Ａユ、、Ａ□２ｔ−
測定しながら制御およびデータ処理コンピュータ１９に
記憶する。以上の動作をくり返しながら反応セ／Ｉ／１
０が反応上／ｌ／１０’の位置にきたところで必要な場
合には第２試薬分注ポンプ２０と連結した第２試薬分注
ノズ／Ｉ／２１が第２試薬分注ノズル移動機構２２によ
って移動し、試薬庫１４内にセットされている第２に薬
容器２３から一定量の＄２試薬２４を吸引し、続いて反
応上／ｌ／１０’のところに移動して反応セ／Ｉ’ＩＯ
内に分注する。第２ＦＳ薬添原後も反応セル１０″が、
洗浄ポンプ２５に連結され、洗浄ノズル上下機構２６に
より上下する洗浄ノズ／ｌ／２７の位置に進む萱での間
も、各位置での吸光度Ａλ□・Ａλ２が測定されコンピ
ュータ１９に記憶されている。制御およびデータ処理コ
ンピュータ１９は各部の動作を同期制御すると同時に９
反応の過程の吸光度データ（各位置におけるＡλ１・Ａ
λ２の値）を用いて、Ｅ料中の目的成分の濃度を算出と
プロゾーン判定を行なう。反応セルは自動洗浄されて再
使用される。また測定項目（目的成分）に応じて、Ｅ料
あるいは標準試料のｇｉｋ変えることができるようにな
っており、また第１試薬あるいは第２試薬も試薬庫内の
別の試薬を使うことができるように構成されている。さ
らに測定項目に応じて測定波長の組合わせも変えること
ができるように構成されている。The spectrometer 17 is moved to the reaction disk 9 by the spectrometer moving mechanism 18.
While reciprocating around the same axis as , for each reaction μ, the absorbance at two wavelengths λ1 and λ2 is calculated as
It is stored in the control and data processing computer 19 as it is being measured. React by repeating the above actions/I/1
When 0 reaches the reaction top/l/10' position, if necessary, the second reagent dispensing nozzle /I/21 connected to the second reagent dispensing pump 20 is moved by the second reagent dispensing nozzle moving mechanism 22. The robot moves to the reaction chamber 14, sucks a certain amount of the $2 reagent 24 from the second reagent container 23 set in the reagent storage 14, and then moves to the reaction chamber /l/10'. IO
Dispense within. Even after the second FS drug addition, the reaction cell 10''
While the cleaning nozzle is connected to the cleaning pump 25 and moved up and down by the cleaning nozzle up/down mechanism 26, the absorbance Aλ□ and Aλ2 at each position is measured and stored in the computer 19. . The control and data processing computer 19 synchronously controls the operation of each part.
Absorbance data during the reaction process (Aλ1・A at each position
λ2 value) is used to calculate the concentration of the target component in the E material and perform prozone determination. The reaction cell is automatically cleaned and reused. In addition, it is possible to change the gik of the E reagent or standard sample depending on the measurement item (target component), and it is also possible to use another reagent in the reagent storage for the first or second reagent. It is composed of Furthermore, the configuration is such that the combination of measurement wavelengths can be changed depending on the measurement item.
また、測定項目に応じて分注する試料量と標準試料量を
独立して変更できる機能を有することがより望ましい・
（へ）発明の効果
この発明によれば、試料に標準試料を添加して試薬と抗
原抗体反応を行なわせることにより容易に超高濃度であ
ること全判定でき、これ萱での簡便的なプロゾーン判定
法で問題魚とされてきた超高濃度試料に対する判定の不
確笑性が解消される。このため、これ壕で確突ではある
が手間や金がかかる方法（希釈険体とのダブノン測定な
ど）全屈いなくとも簡便法を併用することが可能とな９
、自動化学分析装置への適用が容易となる。Furthermore, it is more desirable to have a function that allows the amount of the sample to be dispensed and the amount of the standard sample to be independently changed according to the measurement item. By performing an antigen-antibody reaction with a reagent, it is easy to determine whether the concentration is ultra-high, and this simple prozone determination method eliminates the uncertainty of determining ultra-high concentration samples, which have been regarded as problematic fish. gender is resolved. For this reason, it is possible to use a simple method in conjunction with a method that is reliable but takes time and money (such as measuring Dubnon with a diluted insulating material) without having to completely compromise.9
, it becomes easy to apply to automatic chemical analyzers.
第１図は、この発明の方法の突施に用いる測定装置の一
例の構成説明図、第２図は血清を数段階希釈した試料に
標準試料（コントロール血清ンを添加してＩｇＧ試薬と
反応させた被検液の分光光度計による３４００ｍでの吸
光度と、試料中のＩｇＧ濃度の関係を示す図、第３図は
そのときの３４００ｍ と７０００ｍの吸光度比Ａ３４
０／Ａ７００と試料中のＩｇＧ灘度の関係？示す図、第
４図は血清全数段階希釈しｆＣ試料とＩｇＧ試薬全反応
させた被検液の分光光度計による３４０ ｎｍと７００
ｎｍでのｇ＆光度の検量線である。Fig. 1 is an explanatory diagram of the configuration of an example of a measuring device used for carrying out the method of the present invention, and Fig. 2 shows a sample in which a standard sample (control serum) is added to a sample prepared by diluting serum in several stages and reacted with an IgG reagent. Figure 3 shows the relationship between the absorbance at 3400 m measured by a spectrophotometer of a test liquid and the IgG concentration in the sample. Figure 3 shows the absorbance ratio A34 at 3400 m and 7000 m at that time.
What is the relationship between 0/A700 and the IgG concentration in the sample? The figure shown in Figure 4 shows the spectrophotometer measurements of the test solution obtained by serially diluting the total number of serum and reacting the fC sample with the IgG reagent at 340 nm and 700 nm.
Calibration curve of g&luminosity in nm.
Claims (1)
いは抗原抗体反応による凝集粒子を含有する反応液に光
を照射して、その見かけの吸光度（濁度）を測定し、抗
原あるいは抗体の濃度を測定する方法において、試料に
他の標準試料を添加しこれと試薬を混合したものを被検
液として、該被験液の吸光度を測定し、該吸光度値より
試料中の抗原または抗体濃度の算出と、前記抗原抗体反
応のプロゾーン現象の判定を行うことを特徴とする免疫
反応におけるプロゾーン判定方法。 ２、標準試料と試薬を混合した反応液の吸光度測定値を
超高濃度試料の判定基準として用い、試料に該標準試料
を添加して試薬を混合した被験液の吸光度値が前記基準
値以下であれば超高濃度試料であると判定する第１の判
定方法と、超高濃度試料でないと判定された試料に対し
て、抗原過剰域での反応か抗体過剰域での反応かを判定
する第２のプロゾーン判定方法を併用することを特徴と
する免疫反応におけるプロゾーン判定方法。[Claims] 1. Irradiating light to a reaction solution containing an antigen-antibody conjugate produced by mixing a sample and a reagent or agglomerated particles resulting from an antigen-antibody reaction, and measuring its apparent absorbance (turbidity). However, in the method of measuring the concentration of an antigen or antibody, the absorbance of the test solution is measured by adding another standard sample to the sample and mixing it with a reagent, and then determining the concentration of the sample from the absorbance value. 1. A method for determining prozone in an immune reaction, comprising calculating the concentration of an antigen or antibody, and determining the prozone phenomenon of the antigen-antibody reaction. 2. Use the measured absorbance value of the reaction solution in which the standard sample and reagent are mixed as the criterion for ultra-high concentration samples, and if the absorbance value of the test solution in which the standard sample is added to the sample and the reagent is mixed is below the reference value. If so, the first determination method determines that the sample is an ultra-high concentration sample, and the second determination method determines whether the sample that is determined not to be an ultra-high concentration sample has a reaction in an antigen-excessive range or an antibody-excessive range. A method for determining prozone in an immune reaction, characterized in that the method for determining prozone in item 2 is used in combination.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63239248A JPH0635976B2 (en) | 1988-09-24 | 1988-09-24 | Prozone determination method in immune reaction |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63239248A JPH0635976B2 (en) | 1988-09-24 | 1988-09-24 | Prozone determination method in immune reaction |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0287063A true JPH0287063A (en) | 1990-03-27 |
| JPH0635976B2 JPH0635976B2 (en) | 1994-05-11 |
Family
ID=17041937
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63239248A Expired - Fee Related JPH0635976B2 (en) | 1988-09-24 | 1988-09-24 | Prozone determination method in immune reaction |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0635976B2 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6514770B1 (en) | 1999-07-30 | 2003-02-04 | Mitsubishi Chemical Corporation | Immunoassay |
| WO2017126227A1 (en) * | 2016-01-22 | 2017-07-27 | 株式会社日立ハイテクノロジーズ | Automatic analyzer and standard solution for evaluating scattered light measurement optical system thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102944672B (en) * | 2012-11-16 | 2015-05-20 | 李方和 | Method for qualitatively and quantitatively detecting target substance to be detected in blood serum by utilizing light initiated chemiluminescence immune assay |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS52125623A (en) * | 1976-04-13 | 1977-10-21 | Chugai Pharmaceut Co Ltd | Quantitative determination of antigenic substances and reagent kit for the same |
| JPS60100764A (en) * | 1983-11-07 | 1985-06-04 | Hitachi Ltd | Method for measuring concentration of antigen or antibody |
-
1988
- 1988-09-24 JP JP63239248A patent/JPH0635976B2/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS52125623A (en) * | 1976-04-13 | 1977-10-21 | Chugai Pharmaceut Co Ltd | Quantitative determination of antigenic substances and reagent kit for the same |
| JPS60100764A (en) * | 1983-11-07 | 1985-06-04 | Hitachi Ltd | Method for measuring concentration of antigen or antibody |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6514770B1 (en) | 1999-07-30 | 2003-02-04 | Mitsubishi Chemical Corporation | Immunoassay |
| WO2017126227A1 (en) * | 2016-01-22 | 2017-07-27 | 株式会社日立ハイテクノロジーズ | Automatic analyzer and standard solution for evaluating scattered light measurement optical system thereof |
| JP2017129532A (en) * | 2016-01-22 | 2017-07-27 | 株式会社日立ハイテクノロジーズ | Automatic analyzer and its standard solution for evaluating scattered light measurement optical system |
| US11692929B2 (en) | 2016-01-22 | 2023-07-04 | Hitachi High-Tech Corporation | Automatic analyzer and standard solution for evaluating scattered light measurement optical system thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0635976B2 (en) | 1994-05-11 |
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