JPH0470565A - Latex agglutination test nd reagent used therein - Google Patents
Latex agglutination test nd reagent used thereinInfo
- Publication number
- JPH0470565A JPH0470565A JP18155790A JP18155790A JPH0470565A JP H0470565 A JPH0470565 A JP H0470565A JP 18155790 A JP18155790 A JP 18155790A JP 18155790 A JP18155790 A JP 18155790A JP H0470565 A JPH0470565 A JP H0470565A
- Authority
- JP
- Japan
- Prior art keywords
- latex
- antibody
- specific gravity
- antigen
- sample
- 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
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 18
- 238000012092 latex agglutination test Methods 0.000 title claims description 11
- 239000004816 latex Substances 0.000 claims abstract description 79
- 229920000126 latex Polymers 0.000 claims abstract description 79
- 230000005484 gravity Effects 0.000 claims abstract description 32
- 230000004520 agglutination Effects 0.000 claims abstract description 29
- 238000012360 testing method Methods 0.000 claims abstract description 26
- 210000004369 blood Anatomy 0.000 claims abstract description 25
- 239000008280 blood Substances 0.000 claims abstract description 25
- 239000000427 antigen Substances 0.000 claims abstract description 22
- 102000036639 antigens Human genes 0.000 claims abstract description 22
- 108091007433 antigens Proteins 0.000 claims abstract description 22
- 238000000034 method Methods 0.000 claims abstract description 22
- 210000000601 blood cell Anatomy 0.000 claims abstract description 10
- 210000003743 erythrocyte Anatomy 0.000 claims abstract description 8
- 239000000725 suspension Substances 0.000 claims abstract description 5
- 230000002776 aggregation Effects 0.000 claims description 5
- 238000005054 agglomeration Methods 0.000 claims 1
- 238000010998 test method Methods 0.000 claims 1
- 210000002966 serum Anatomy 0.000 abstract description 9
- 102000004190 Enzymes Human genes 0.000 abstract description 3
- 108090000790 Enzymes Proteins 0.000 abstract description 3
- 239000011248 coating agent Substances 0.000 abstract description 3
- 238000000576 coating method Methods 0.000 abstract description 3
- 239000005556 hormone Substances 0.000 abstract description 3
- 229940088597 hormone Drugs 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 3
- 239000002245 particle Substances 0.000 description 10
- 239000000126 substance Substances 0.000 description 9
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 7
- 239000000047 product Substances 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- OOIBFPKQHULHSQ-UHFFFAOYSA-N (3-hydroxy-1-adamantyl) 2-methylprop-2-enoate Chemical compound C1C(C2)CC3CC2(O)CC1(OC(=O)C(=C)C)C3 OOIBFPKQHULHSQ-UHFFFAOYSA-N 0.000 description 4
- 239000004471 Glycine Substances 0.000 description 4
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 4
- 238000004220 aggregation Methods 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 241000283973 Oryctolagus cuniculus Species 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 description 2
- 230000009260 cross reactivity Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000009169 immunotherapy Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 230000007721 medicinal effect Effects 0.000 description 1
- 238000009304 pastoral farming Methods 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000186 progesterone Substances 0.000 description 1
- 229960003387 progesterone Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
【発明の詳細な説明】
［産業上の利用分野］
本発明は、ラテックス凝集試験方法に関し、さらに詳細
には、血液中に存在するホルモン、酵素、タンパク質等
や抗体等を測定するに当り、血清分離を必要としないラ
テックス凝集試験方法に関する。[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a latex agglutination test method, and more specifically, in measuring hormones, enzymes, proteins, etc., antibodies, etc. present in blood, serum This invention relates to a latex agglutination test method that does not require separation.
［従来の技術及びその課題］
ラテックス凝集試験は、ラテックス粒子を被検物質に対
する抗体あるいは抗原で被覆し、被検試料、例えば、血
清、尿、培養液、抽出液等と混合、接触させ、生じたラ
テックス粒子の凝集を肉眼で判断することにより被検試
料中の抗体あるいは抗原量（被検物質量）を測定する方
法である。[Prior art and its problems] In the latex agglutination test, latex particles are coated with antibodies or antigens against the test substance, mixed with a test sample such as serum, urine, culture solution, extract, etc., and brought into contact with the latex particles. This method measures the amount of antibodies or antigens (amount of test substance) in a test sample by visually determining the aggregation of latex particles.
しかしながら、従来ラテックス凝集試験法において全血
は被検試料として用いることはできないとされており、
また、全血を試料として用いた試験結果が報告された例
もない。However, it has been said that whole blood cannot be used as a test sample in the conventional latex agglutination test method.
Furthermore, there are no reports of test results using whole blood as a sample.
その理由は、■全血を試料とした場合、ラテックスの凝
集塊が赤血球の下に沈み、その生成が確認できない、■
赤血球の下に沈み込んだ凝集塊を観察しようとして、例
えば試験平板を傾けても自然沈澱のものと凝集によるも
のとが判別できない点にあっ力。The reason for this is: ■When whole blood is used as a sample, latex aggregates sink below the red blood cells, making it impossible to confirm their formation.■
I was impressed by the fact that even if I tried to observe the aggregates that had sunk beneath the red blood cells, for example, by tilting the test plate, I could not distinguish between natural sedimentation and agglutination.
このような理由により、全血を試験試料として使用する
ことはなく、血清を利用しているのであるが、血清を得
るためには、血液を遠心分離等にかけなければならず、
例えばこのような装置のない屋外において緊急に試験を
行う必要がある場合にはラテックス凝集法は使用するこ
とはできなかった。For these reasons, whole blood is not used as a test sample, but serum is used; however, in order to obtain serum, blood must be subjected to centrifugation, etc.
For example, the latex agglutination method could not be used when there was an urgent need to conduct a test outdoors where such equipment was not available.
［課題を解決するための手段］
本発明者らは、ラテックス凝集法の適用を全血試料に対
しても広げるべく鋭意研究をおこなった結果、ラテック
ス粒子の比重を適切に選択し、反応により生じたラテッ
クス凝集塊が反応系の表面に浮上するようにすれば、全
血試料に対してもラテックス凝集法が適用し得ることを
見いだした。[Means for Solving the Problems] The present inventors conducted intensive research to extend the application of latex agglutination method to whole blood samples, and as a result, the specific gravity of latex particles was appropriately selected, and the We have discovered that the latex agglutination method can also be applied to whole blood samples by allowing the latex aggregates to float to the surface of the reaction system.
すなわち本発明は、試料中の抗原または抗体と、これに
対する抗体または抗原を結合したラテックスを反応させ
、生じたラテックス凝集により試料中の抗原または抗体
の存在を測定するラテックス凝集試験方法において、試
験試料として赤血球を含む全血試料を用い、反応により
生じたラテックス凝集を反応溶液表面上に浮上させるこ
とを特徴とするラテックス凝集試験方法を提供するもの
である。That is, the present invention provides a latex agglutination test method in which an antigen or antibody in a sample is reacted with latex to which the antibody or antigen is bound, and the presence of the antigen or antibody in the sample is measured by the resulting latex agglutination. The present invention provides a latex agglutination test method characterized in that a whole blood sample containing red blood cells is used as a sample, and latex agglutination generated by the reaction is floated on the surface of a reaction solution.
本発明方法においては、ラテックス凝集を反応溶液表面
に浮上させるため、試験に使用する抗体または抗原を結
合したラテックス（以下、 「試験ラテックス」という
）の比重及び反応系の液性、例えば塩類やタンパク質等
の種類及び濃度が重要である。 具体的には、試験ラテ
ックスの比重が全血試料とラテックス懸濁液からなる反
応系の非血球画分の比重とほぼ同一かまたはわずかに高
く、がっ、血球画分の比重より低いことが必要である。In the method of the present invention, in order to float latex aggregates to the surface of the reaction solution, the specific gravity of the latex bound to the antibody or antigen used in the test (hereinafter referred to as "test latex") and the liquid properties of the reaction system, such as salts and protein, are determined. The type and concentration of the substances are important. Specifically, the specific gravity of the test latex is approximately the same or slightly higher than the specific gravity of the non-blood cell fraction of the reaction system consisting of a whole blood sample and a latex suspension, but it is lower than the specific gravity of the blood cell fraction. is necessary.
このような試験ラテックスを調製するためには、原料ラ
テックスとしてその比重が０．９９０−１．０８０程度
の低比重ラテックスを使用し、抗体または抗原での被覆
を上記条件の比重範囲に入るようにすれば良い。なお、
この試験ラテックスの好ましい比重範囲は、測定系の比
重等によって異なってくるので数字で限定することはで
きないが、−Ｇには１．０１５〜１．０８０程度である
。In order to prepare such a test latex, a low specific gravity latex with a specific gravity of about 0.990-1.080 is used as the raw material latex, and the coating with the antibody or antigen is adjusted so that the specific gravity falls within the specific gravity range of the above conditions. Just do it. In addition,
The preferred specific gravity range of this test latex varies depending on the specific gravity of the measuring system, etc., and therefore cannot be limited numerically, but for -G it is about 1.015 to 1.080.
抗体、抗原での原料ラテックスの被覆は、常法にしたが
って行えばよく、特に制限はない。Coating of the raw material latex with antibodies and antigens may be carried out according to a conventional method, and there are no particular limitations.
本発明は、後でも示すように、試験ラテックスとこれが
凝集したラテックスの比重、反応系の液性および血球画
分の比重が重要であ本発明のラテックス凝集試験におい
ては、被測定物質により、添加される試薬、塩類及び蛋
白濃度が異なるので、使用すべき試験ラテックス及び凝
集したラテックスの比重や反応系の液性を試験的に定め
ることが必要である。As will be shown later, in the latex agglutination test of the present invention, the specific gravity of the test latex and its agglomerated latex, the humoral nature of the reaction system, and the specific gravity of the blood cell fraction are important. Since the reagents, salts, and protein concentrations used differ, it is necessary to experimentally determine the specific gravity of the test latex and aggregated latex to be used, as well as the liquid properties of the reaction system.
そして、試験ラテックスの比重の調製は、原料ラテック
スの比重と原料ラテックスに対する抗原または抗体の被
覆量を調製することにより行うことができる。The specific gravity of the test latex can be adjusted by adjusting the specific gravity of the raw latex and the amount of antigen or antibody coated on the raw latex.
本発明は、試験に用いるラテックスとして上記した試験
ラテックスを用い、被検試料として全血試料を用いる以
外は、公知のラテックス凝集法と同様実施することがで
きる。The present invention can be carried out in the same manner as known latex agglutination methods, except that the above-mentioned test latex is used as the latex used in the test and a whole blood sample is used as the test sample.
すなわち、全血試料に試験ラテックスを添加し、反応さ
せた後、反応系表面に浮上してくる凝集ラテックスから
被測定物質の存在を判断すれば良い。That is, after adding a test latex to a whole blood sample and allowing it to react, the presence of the substance to be measured may be determined from the aggregated latex that floats to the surface of the reaction system.
［作 用 ］
ラテックス凝集反応は、ラテックス粒子表面に抗原ある
いは抗体を被覆し、その抗原あるいは抗体に反応性のあ
る抗体あるいは抗原をそのラテックスの凝集により測定
するものである。 本発明は、種々の反応液性における
凝集用ラテックス粒子の比重が血球成分の比重に比べ低
く、血清成分よりも高いか同等の比重を有することに着
目した反応系である。[Function] In the latex agglutination reaction, the surface of latex particles is coated with an antigen or antibody, and antibodies or antigens that are reactive with the antigen or antibody are measured by agglutination of the latex. The present invention is a reaction system that focuses on the fact that the specific gravity of latex particles for aggregation in various reaction liquids is lower than that of blood cell components, and higher than or equal to serum components.
すなわち本願発明の凝集試薬は、全血試料に加えられた
場合、均一な反応系を形成し、反応により生じたラテッ
クス凝集塊は浮力により反応溶液表面上に浮遊し、陰性
と陽性との判定を可能とするという新しい技術思想に基
づくものである。That is, when the agglutination reagent of the present invention is added to a whole blood sample, it forms a homogeneous reaction system, and the latex aggregates generated by the reaction float on the surface of the reaction solution due to buoyancy, making it possible to determine whether the sample is negative or positive. It is based on a new technological idea that makes it possible.
［発明の効果］
従来ラテックス凝集法は、全血試料に対しては適用する
ことができず、血清分離を行って初めて実施することが
できた。 しかし本発明方法によれば、血清分離するこ
となく、血液中に存在するホルモン、酵素、抗体等を検
出することが可能になった。[Effects of the Invention] Conventional latex agglutination methods could not be applied to whole blood samples, and could only be performed after serum separation. However, according to the method of the present invention, it has become possible to detect hormones, enzymes, antibodies, etc. present in blood without separating serum.
従って、本発明によれば血液中の上記成分の検圧が容易
になったのみならず、例えば、畜産等の現場で血清分離
が困難な状況であっても、例えば、授精卵移植のときに
必要な、血中のプロゲステロン量を測定を迅速に行うこ
とが可能となり、極めて有利に使用できる。Therefore, according to the present invention, it is not only easy to measure the pressure of the above-mentioned components in blood, but also, even in situations where serum separation is difficult in livestock farming, etc., for example, when performing fertilized egg transplantation. It becomes possible to quickly measure the necessary amount of progesterone in the blood, and it can be used extremely advantageously.
［実施例 ］
以下実施例をもって本発明をさらに詳しく説明する。
以下の実Ｆｊ、ｆＮでは、マウスモノクローナル抗体に
対する抗体（以下、 ｒＨＡＭＡ」という）について本
発明方法で試験した結果を示すが、本発明方法は当然の
ことながらこれらの物質に適用が限定されるものではな
い。[Example] The present invention will be explained in more detail with reference to Examples below.
The following examples Fj and fN show the results of testing antibodies against mouse monoclonal antibodies (hereinafter referred to as "rHAMA") using the method of the present invention; however, the method of the present invention is naturally limited in its applicability to these substances. isn't it.
なお、実施例におけるＨＡＭＡおよびＨＡＭＡ様物Ｎ（
マウスモノクローナル抗体と交差反応性を示す物質）の
測定は、マウスモノクローナル抗体を利用するイムノシ
ンチおよびイムノセラフイーにおいて重要な意義のある
ものである。In addition, HAMA and HAMA-like substance N (
The measurement of substances that exhibit cross-reactivity with mouse monoclonal antibodies is of important significance in immunoscintigraphy and immunotherapy that utilize mouse monoclonal antibodies.
すなわち、イムノシンチやイムノセラフイーは同一人に
対し、複数回適用される医療技術であるが、マウスモノ
クローナル抗体を人体に適用した場合に、これに対する
抗体（ＨＡＭＡ）が産生され、この結果、イムノコンプ
レックスが生じ本来の薬剤投与の効果を期待てきなくな
る。In other words, immunoscintigraphy and immunotherapy are medical techniques that are applied multiple times to the same person, but when a mouse monoclonal antibody is applied to the human body, antibodies against it (HAMA) are produced, and as a result, immunocomplex This occurs and the intended effect of drug administration is no longer expected.
また、人体中に一部のリュウマチ因子や、扶植（ｓｓＤ
ＮＡ）抗体が存在するとこれがマウスモノクローナル抗
体と交差反応性を示しＨＡＭＡ様物質同様に本来の医療
効果を期待できなくなる。In addition, some rheumatoid factors and ssD are present in the human body.
NA) If antibodies are present, they will show cross-reactivity with mouse monoclonal antibodies, and similar to HAMA-like substances, original medical effects cannot be expected.
従って、イムノシンチ剤やイムノセラフイー剤を投与す
る前に、体内に存在するＨＡＭＡおよびＨＡＭＡ様物質
置物質量することは医療技術上重要である。Therefore, it is important in medical technology to determine the amount of HAMA and HAMA-like substances present in the body before administering immunoscinticulture agents or immunotherapeutic agents.
実施例 １
低比重ラテックス凝集試薬（本願発明
品）の調製：
５０ｍ１容遠心管にラテックス浮遊液（＃Ｉ水化学工業
（株）製Ｎ−１０００（粒径１、ＯＯミ７［１：／
固形分１（Ｈ比重１．００３） １ ｍ ｌと０．０
５Ｍグリシン緩衝液（ｐＨ８，５，０，１５Ｍ塩化ナト
リウムを含む）９ｍｌに、マウスモノクローナル抗体を
１ｍｇ／ｍ］となるように加えた。 この混合物をポ
ルテックスにて撹拌後、５０ ’Ｃにて３０分間靜装し
た。 高速遠心機（ＫＵＢＯＴＡ ＲＲ／２００００
）にて４０００回転／分で１５分間遠心した。 上澄み
を除去後、０．０１Ｍグリシン緩衝液（ｐＨ８，６、１
％ＢＳＡ、０．１５Ｍ塩化ナトリウムを含む）１０ｍｌ
にてラテックスを再浮遊させた。 パラフィルムにて遠
心管の口を閉じ、４℃にて一晩装置シタ。４０００回転
／分で１５分間遠心し、上澄みを除去した。 ０．０５
Ｍグリシン緩衝液（ｐＨ８，６，０，０５％ＢＳＡ、
０．０１％アジ化ナトリウム、０．１５Ｍ塩化ナトリウ
ムを含む） １０ｍ１にてラテックスを再浮遊させた
。 ４０００回転／分で１５分間遠心し、上澄みを除
去した。 この操作を３回ｍ逐した。 ０．０５ Ｍグ
リシン緩衝液（ｐＨ８，６，０，０５％ＢＳＡ、０．０
１％アジ化ナトリウム、０．１５Ｍ塩化ナトリウムを含
む）５ｍｌに再浮遊させ、低比重ラテックス凝集試薬（
ラテックス分２％）溶液を得た。Example 1 Preparation of low specific gravity latex agglutination reagent (product of the present invention): In a 50 ml centrifuge tube, a latex suspension (#I N-1000 manufactured by Mizu Kagaku Kogyo Co., Ltd. (particle size 1, OO Mi 7 [1:/
Solid content 1 (H specific gravity 1.003) 1 ml and 0.0
A mouse monoclonal antibody was added to 9 ml of 5M glycine buffer (pH 8, 5, 0, containing 15M sodium chloride) at a concentration of 1 mg/m. This mixture was stirred with a portex and then kept at 50'C for 30 minutes. High-speed centrifuge (KUBOTA RR/20000
) for 15 minutes at 4000 rpm. After removing the supernatant, add 0.01M glycine buffer (pH 8, 6, 1
%BSA, 0.15M sodium chloride) 10ml
The latex was resuspended. Close the centrifuge tube with parafilm and leave the apparatus at 4°C overnight. It was centrifuged at 4000 rpm for 15 minutes and the supernatant was removed. 0.05
M glycine buffer (pH 8, 6, 0, 05% BSA,
The latex was resuspended in 10ml (containing 0.01% sodium azide, 0.15M sodium chloride). It was centrifuged at 4000 rpm for 15 minutes and the supernatant was removed. This operation was repeated three times. 0.05 M glycine buffer (pH 8,6, 0.05% BSA, 0.0
resuspended in 5 ml of low density latex agglutination reagent (containing 1% sodium azide, 0.15M sodium chloride) and
A solution with a latex content of 2% was obtained.
比較例
ラテックス浮遊液として、高比重ラテックス（武田薬品
工業（株）ＷＳＤＬ−５９粒径０．９０ミクａｙ 固
形分５χ 比重１．１２） ２ ｍ ｌを用いる以外は
実施例１と同様にして高比重ラテックス凝集試薬（ラテ
ックス分２％）溶液（比較品）を得た。Comparative Example A high-density latex (Takeda Pharmaceutical Co., Ltd. WSDL-59 particle size 0.90 mAy, solid content 5χ, specific gravity 1.12) was used in the same manner as in Example 1, except that 2 ml of high-density latex was used as the latex suspension. A specific gravity latex agglutination reagent (latex content 2%) solution (comparative product) was obtained.
実施例２
低比重ラテックス凝集試薬と高比重
ラテックス凝集試薬の比較：
実施例１と比較例で調製したラテックス凝集試薬をもち
い、以下に示す方法によりＨＡＭＡの検出について試験
した。 この結果を第１表に示す。Example 2 Comparison of low-density latex agglutination reagent and high-density latex agglutination reagent: Using the latex agglutination reagent prepared in Example 1 and Comparative Example, HAMA detection was tested by the method shown below. The results are shown in Table 1.
（１）測定方法
ラテックス凝集判定プレート（覆水化学工業（株）製
Ｃ−ＨＰＩＯ）のＮ、１．２及び３の反応区域枠内に低
比重ラテックス凝集試薬（本発明品）あるいは高比重ラ
テックス凝集試薬（比較品）をそれぞれ５０μｌずつ滴
下する。 Ｎ０８１の反応区域枠内には、正常兎よりヘ
パリン採血した新鮮血を用いてアフィニティーｍ製され
た抗マウス エｇＧ（Ｈ＋Ｌ）ウサギ抗体（カッペル社
製１０６１１−００８２）を１０００倍希釈したもの５
０μｍを加え撹拌した。 Ｎ ｏ 、 ２の反応区域枠
内には、正常兎よりヘパリン採血した新鮮血を５０μｍ
滴下し、撹拌棒で混ぜ合わせ、反応区域枠内に広げた。(1) Measuring method Latex agglutination determination plate (manufactured by Okisui Kagaku Kogyo Co., Ltd.)
Drop 50 μl each of a low-density latex agglutination reagent (product of the present invention) or a high-density latex agglutination reagent (comparative product) into the reaction zone frames of N, 1.2, and 3 of C-HPIO). In the reaction area frame of N081, anti-mouse egg G (H+L) rabbit antibody (manufactured by Kappel Co., Ltd. 10611-0082) prepared by affinity m using fresh heparinized blood from a normal rabbit was diluted 1000 times.
0 μm was added and stirred. Within the reaction area frame of No. 2, fresh blood collected from a normal rabbit with heparin was collected at a thickness of 50 μm.
Dropwise, mix with a stir bar and spread within the reaction zone frame.
Ｎ ｏ 、 ３の反応区域枠内には、Ｎ ｏ 、 １
に加えた試料と４．３ｍｇ／ｍｌのマウスモノクローナ
ル抗体溶液（０，ＯＩＭリン酸緩１ｉｉｉ液）２５μｍ
を滴下し、撹拌棒で混ぜ合わせ、反応区域枠内に広げた
。 それぞれ１分間ゆるやかに判定板を動かし、１０分
間靜１し、凝集像を観察し判定した。Within the reaction zone frame of N o , 3, there are N o , 1
sample and 4.3 mg/ml mouse monoclonal antibody solution (0, OIM phosphate 1III solution) 25 μm
was added dropwise, mixed with a stir bar, and spread within the reaction zone frame. The evaluation plate was moved gently for 1 minute each, and then kept still for 10 minutes, and the agglutination image was observed and evaluated.
（２）測定結果
第１表
Ｎｏ、Ｉ Ｎｏ、２ Ｎｏ、３低比重ラテ
ックス凝集試薬 陽 性 陰 性 陰 性（
本発明品）
高比重ラテックス凝集試薬 判定不可 判定不可
判定不可（比較品）
これに比べ、低比重ラテックス凝集試薬のＮｏ、２と３
は未凝集のラテックス粒子が赤血球の上に未凝集の状態
で確認でき、陰性判定も容易に出来た。(2) Measurement results Table 1 No. I No. 2 No. 3 Low specific gravity latex agglutination reagent Positive Negative Negative (
Product of the present invention) High-density latex agglutination reagent Unable to determine Unable to determine
Undeterminable (comparative product) In comparison, low-density latex agglutination reagents No. 2 and 3
In this case, unagglutinated latex particles were confirmed on red blood cells in an unagglutinated state, and a negative judgment was easily made.
なお、上記の各状態を示す写真を参考写真として示す。Note that photographs showing each of the above conditions are shown as reference photographs.
以 上
二の結果から明らかなように、低比重ラテックス凝集試
薬を用いた場合、Ｎｏ、１のラテックス凝集塊は反応溶
液表面上に浮かび上がり、凝集が明瞭に判定できた。
これに対し、高比重ラテックス試薬のＮｏ、１の凝集塊
は赤血球の下に沈み込及 その凝集塊を確認できなかっ
た。 そればかりか１０分間靜１しておくことによりラ
テックス粒子は赤血球の下に沈み込み、Ｎｏ、２と３の
判定が不可能であっ出願人 株式会社第一ラジオアイソAs is clear from the above two results, when the low specific gravity latex aggregation reagent was used, latex aggregates No. 1 floated on the surface of the reaction solution, and aggregation could be clearly determined.
On the other hand, the aggregates of high-density latex reagent No. 1 sank below the red blood cells, and the aggregates could not be confirmed. Not only that, but by leaving it still for 10 minutes, the latex particles sink under the red blood cells, making it impossible to determine No. 2 and No. 3. Applicant: Daiichi Radioiso Co., Ltd.
Claims (3)
たは抗原を結合したラテックスを反応させ、生じたラテ
ックス凝集により試料中の抗原または抗体の存在を測定
するラテックス凝集試験方法において、 試験試料として赤血球を含む全血試料を用い、反応によ
り生じたラテックス凝集を反応溶液表面上に浮上させる
ことを特徴とするラテックス凝集試験方法。(1) In a latex agglutination test method in which an antigen or antibody in a sample is reacted with latex bound to the antibody or antigen, and the presence of the antigen or antibody in the sample is measured by the resulting latex agglutination, red blood cells are used as the test sample. 1. A latex agglutination test method, which uses a whole blood sample containing .
血試料とラテックス懸濁液からなる反応系の非血球画分
の比重とほぼ同一かまたはわずかに高く、かつ、血球画
分の比重より低いことを特徴とする請求項第１項記載の
ラテックス凝集試験方法。(2) The specific gravity of the latex bound to the antibody or antigen is approximately the same or slightly higher than the specific gravity of the non-blood cell fraction in the reaction system consisting of a whole blood sample and a latex suspension, and is lower than the specific gravity of the blood cell fraction. The latex agglomeration test method according to claim 1, characterized in that:
とほぼ同一かまたはわずかに高く、かつ、血球画分の比
重より低くなるように、被検試料に対する抗体または抗
原を結合させたラテックスを含有するラテックス凝集試
験用試薬。(3) Bind the antibody or antigen to the test sample so that the specific gravity is approximately the same or slightly higher than the specific gravity of the non-blood cell fraction in the reaction system including the whole blood sample, and lower than the specific gravity of the blood cell fraction. A latex agglutination test reagent containing a latex that has been
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18155790A JP2895584B2 (en) | 1990-07-11 | 1990-07-11 | Latex agglutination test method and reagents used therefor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18155790A JP2895584B2 (en) | 1990-07-11 | 1990-07-11 | Latex agglutination test method and reagents used therefor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0470565A true JPH0470565A (en) | 1992-03-05 |
| JP2895584B2 JP2895584B2 (en) | 1999-05-24 |
Family
ID=16102871
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP18155790A Expired - Lifetime JP2895584B2 (en) | 1990-07-11 | 1990-07-11 | Latex agglutination test method and reagents used therefor |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2895584B2 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9632537B2 (en) | 2013-09-23 | 2017-04-25 | Apple Inc. | Electronic component embedded in ceramic material |
| US9678540B2 (en) | 2013-09-23 | 2017-06-13 | Apple Inc. | Electronic component embedded in ceramic material |
-
1990
- 1990-07-11 JP JP18155790A patent/JP2895584B2/en not_active Expired - Lifetime
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9632537B2 (en) | 2013-09-23 | 2017-04-25 | Apple Inc. | Electronic component embedded in ceramic material |
| US9678540B2 (en) | 2013-09-23 | 2017-06-13 | Apple Inc. | Electronic component embedded in ceramic material |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2895584B2 (en) | 1999-05-24 |
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