JP2587969B2 - Virus antibody detection reagent - Google Patents

Virus antibody detection reagent

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Publication number
JP2587969B2
JP2587969B2 JP62326774A JP32677487A JP2587969B2 JP 2587969 B2 JP2587969 B2 JP 2587969B2 JP 62326774 A JP62326774 A JP 62326774A JP 32677487 A JP32677487 A JP 32677487A JP 2587969 B2 JP2587969 B2 JP 2587969B2
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Japan
Prior art keywords
hiv
protein
antibody
gelatin particles
env
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Japanese (ja)
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JPH01169360A (en
Inventor
克仁 川戸
幹雄 池田
隆一 藤野
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Fujirebio Inc
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Fujirebio Inc
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Description

【発明の詳細な説明】 [産業上の利用分野] 本発明は後天性免疫不全症候群関連ウイルス抗体検出
用試薬に関し、さらに詳しくは、遺伝子組み換えHIV抗
原を用いた後天性免疫不全症候群関連ウイルス抗体検出
用間接凝集反応試薬に関する。
Description: TECHNICAL FIELD The present invention relates to a reagent for detecting an acquired immunodeficiency syndrome-associated virus antibody, and more particularly to detection of an acquired immunodeficiency syndrome-associated virus antibody using a recombinant HIV antigen. For indirect agglutination reaction reagents.

[従来の技術] 後天性免疫不全症候群(Acquired Immunodeficiency
Syndrome−通常AIDSと呼ばれている)は、1981年に初め
て認められた新しい病気である。[M.S.Gottilb et a
l.,N.Eng,J.Med.,305、1425(1981)]。その後、フラ
ンスでリンパ節疾患症候群(Lymphadenopathy Syndrom
e)患者からLAVとなずけられたレトロウイルスが[F.Ba
rrs−Sinoussi et al.,Science、220、868(1983)]、
そして、アメリカでAIDS患者やpre−AIDS患者からTHLV
−III[M.Popovic etal.,Science、224、497(1984)]
及びARV[J.A.Levy et al.,Science、225、840(198
4)]となずけられたレトロウイルスが相次いで発見さ
れた。最近、これらのLAV、HTLV−III及びARVはその遺
伝子の塩基配列が完全に解析され、これらのレトロウイ
ルスは同じウイルスの変異株であることが明らかになっ
た[L.Ratner et al.,Nature、313、277(1985)、R.Sa
nchez Pescaoor et al.,Science、227、484(1985),S.
Wain Hobson et al.,Cell,40,(1985)]。
[Related Art] Acquired Immunodeficiency Syndrome
Syndrome (commonly called AIDS) is a new disease first recognized in 1981. [MSGottilb et a
l., N. Eng, J. Med., 305, 1425 (1981)]. Later, in France, lymph node disease syndrome (Lymphadenopathy Syndrom
e) A retrovirus identified as LAV by the patient is [F.
rrs-Sinoussi et al., Science, 220, 868 (1983)],
And in the United States, AILV patients and pre-AIDS patients
−III [M. Popovic et al., Science, 224, 497 (1984)]
And ARV [JA Levy et al., Science, 225, 840 (198
4)] were discovered one after another. Recently, the LAV, HTLV-III, and ARV genes have been completely sequenced and their retroviruses have been shown to be mutants of the same virus [L. Ratner et al., Nature. , 313, 277 (1985), R. Sa
nchez Pescaoor et al., Science, 227, 484 (1985), S.
Wain Hobson et al., Cell, 40, (1985)].

このAIDSの病因ウイルスは一般にHIV(Human Immunod
eficiency Virus)と総称されているが、HIVはヒトとヒ
トとの接触、輸血等により感染し、発病する。この際、
感染したヒトの血液中にAIDS関連ウイルスと特異的に反
応する抗体が生ずることも知られている。従ってこの抗
体を検出することによって、HIV感染の有無を調べれ
ば、AIDSの予防につながるものと考えられる。
This AIDS-causing virus is generally HIV (Human Immunod
HIV is commonly referred to as eficiency virus), but it is transmitted and infected by human-to-human contact and blood transfusion. On this occasion,
It is also known that antibodies reacting specifically with AIDS-related viruses occur in the blood of infected humans. Therefore, if the presence or absence of HIV infection is detected by detecting this antibody, it is considered that this will lead to the prevention of AIDS.

そこで、本発明者らは先に、HIVの抗原成分を感作し
た担体粒子からなるAIDS関連ウイルス抗体検出用間接凝
集反応試薬を提案した(特開昭62−182662号公報参
照)。
Accordingly, the present inventors have previously proposed an indirect agglutination reagent for detecting an AIDS-related virus antibody comprising carrier particles sensitized with an HIV antigen component (see JP-A-62-182662).

一方、HIVはRNAを核として持ち、これと逆転写酵素を
包む核タンパク(coreタンパク)と、さらにこれを包み
込む外殻糖タンパク(envタンパク)から構成されてい
る。
On the other hand, HIV has RNA as a nucleus, and is composed of a nuclear protein (core protein) that wraps the RNA and a reverse transcriptase, and an outer glycoprotein (env protein) that further wraps the nucleus.

HIVの感染によって生体内ではこれらウイルスの構成
タンパクに対する抗体が産生されるが、これまでの研究
によれば、HIVには感染しているが発病していないいわ
ゆるキヤリア群とAIDSを発病した群との間で、p24に代
表されるcoreタンパクとgp41に代表されるenvタンパク
のそれぞれのタンパクに対する抗体量に差異が見られた
という報告がいくつか発表されている。そしてHIVへの
感染により通常coreタンパク及びenvタンパクの両方に
対する抗体が産生されるが、感染後ある程度の期間が経
過すると、それらの産生量に差異が生じ、coreタンパク
に対する抗体価が低下しはじめる頃からARC(AIDS rela
bed couplex)の症状がではじめ、AIDSへと移行してい
くとの報告もされている(Joepet al.,British Med.J.,
1986)。
HIV infection produces antibodies against the constituent proteins of these viruses in vivo.According to previous studies, there are so-called carrier groups that have been infected but have not developed HIV and those that have developed AIDS. Some reports have shown that differences were found in the amount of antibody against each of the core protein represented by p24 and the env protein represented by gp41. HIV infection usually produces antibodies against both the core protein and the env protein.However, after a certain period of time after infection, a difference in the amount of their production occurs and the antibody titer against the core protein begins to decrease. From ARC (AIDS rela
It has been reported that the symptoms of bed couplex begin to shift to AIDS (Joepet al., British Med. J.,
1986).

従って、抗HIV−core(p24)クンパク抗体及び抗HIV
−env(gp41あるいはp41)タンパク抗体の検出はHIVへ
の感染からキヤリアを経てARC、AIDS発症へと移行する
過程を追跡する上で臨床的意義が極めて大きいといえ
る。
Therefore, anti-HIV-core (p24) anti-HIV antibody and anti-HIV
The detection of -env (gp41 or p41) protein antibody is of great clinical significance in tracking the transition from HIV infection to ARC and AIDS through carriers.

従来よりこれら抗HIV−core(p24)タンパク抗体、HI
V−env(gp41あるいはp41)タンパク抗体をウエスター
ン・ブロット法(Western blot法)又はEIA法で測定す
ることが提案されている[LANCET、1986年11月29日号12
33〜1237頁参照]。しかし、前者のウエスターン・ブロ
ット法はHIV抗原タンパク成分をSDS電気泳動後ニトロセ
ルロース膜に転写し、この膜を被検血清とEIA法やRIA法
で反応させるものであるが、操作が煩雑で結果が出るま
でに2日以上もの時間がかかるという欠点がある。また
後者のEIA法(Abbot社法)においても、検体との反応に
十数時間を要し、反応の工程数も多く手段的にも煩雑で
ある。
Conventionally, these anti-HIV-core (p24) protein antibodies, HI
It has been proposed to measure V-env (gp41 or p41) protein antibody by Western blotting or EIA [LANCET, Nov. 29, 1986, December 12, 1986].
See pages 33 to 1237]. However, in the former Western blot method, the HIV antigen protein component is transferred to a nitrocellulose membrane after SDS electrophoresis, and this membrane is reacted with the test serum by the EIA or RIA method, but the operation is complicated. The disadvantage is that it takes more than two days for the results to be obtained. Also, in the latter EIA method (Abbot's method), the reaction with the sample requires ten and several hours, the number of reaction steps is large, and the method is complicated.

[発明が解決しようとする問題点] 本発明は、上記従来法における欠点を克服し、簡便で
迅速に抗HIV−core(p24)タンパク抗体及び抗HIV−env
(p41)タンパク抗体を検出することのできる検出用試
薬を提供することを目的とするものであり、本発明者ら
はこれを目的に鋭意検討を行った結果、今回、HIV−cor
e(p24)タンパク及び/又はHIV−env(p41)タンパク
を成る特定のpH条件下で感作した担体粒子を用いると、
間接凝集反応により、簡便かつ迅速に検体中の抗HIV−c
ore(p24)タンパク抗体、抗HIV−env(p41)タンパク
抗体を検出しうることを見い出し本発明を完成するに至
った。
[Problems to be Solved by the Invention] The present invention overcomes the above-mentioned drawbacks of the conventional method, and provides a simple and rapid anti-HIV-core (p24) protein antibody and anti-HIV-env.
(P41) It is an object of the present invention to provide a detection reagent capable of detecting a protein antibody, and the present inventors have conducted intensive studies for this purpose.
Using carrier particles sensitized with e (p24) protein and / or HIV-env (p41) protein under specific pH conditions,
Simple and quick anti-HIV-c
The inventors have found that an ore (p24) protein antibody and an anti-HIV-env (p41) protein antibody can be detected, and have completed the present invention.

[問題点を解決するための手段] しかして、本発明によれば、HIV−p24及び/又はHIV
−p41をp7.3〜8.5において感作いたゼラチン粒子からな
ることを特徴とする後天性免疫不全症候群(AIDS)関連
ウイルス抗体検出用間接凝集反応試薬が提供される。
[Means for Solving the Problems] However, according to the present invention, HIV-p24 and / or HIV
-An indirect agglutination reagent for detecting an acquired immunodeficiency syndrome (AIDS) -related virus antibody characterized by comprising gelatin particles sensitized to p41 at p7.3 to 8.5.

HIV−p24はHIVの核に存在するRNAをとり囲む分子量が
約2400のcoreタンパクの一種であり、またHIV−p41はHI
Vの包被を構成する糖タンパクの一種であるgp−41(分
子量約4100)から糖を除いた成分であり、これらタンパ
クはいずれも既知であり、例えば、ANALYTICAL BIOCHE
MISTRY 1613、370〜379(1987)に記載の方法に従い、
大腸菌等の菌を用いた遺伝子組換え法によりHIV−p24及
びHIV−p41を得ることができる。これらHIV−p24及びHI
V−p41はそれぞれ単独で用いてもよく、或いは両者を併
用してもよい。
HIV-p24 is a kind of core protein with a molecular weight of about 2400 surrounding RNA present in the nucleus of HIV, and HIV-p41 is HIV-p41.
V is a component obtained by removing sugar from gp-41 (a molecular weight of about 4100), which is a kind of glycoprotein constituting the envelope of V. All of these proteins are known. For example, ANALYTICAL BIOCHE
According to the method described in MISTRY 1613 , 370-379 (1987),
HIV-p24 and HIV-p41 can be obtained by a gene recombination method using a bacterium such as Escherichia coli. These HIV-p24 and HI
V-p41 may be used alone or in combination.

上記HIV−p24及び/又はHIV−p41で感作するためのゼ
ラチン粒子(この粒子の製造法について特開昭57−1536
58号公報参照)は非特異反応が少なく、用途に応じた性
質を付加できる等、AIDS関連ウイルス抗体検出のための
間接凝集反応用担体として優れた長所を有している。
Gelatin particles for sensitization with the above-mentioned HIV-p24 and / or HIV-p41 (Japanese Patent Application Laid-Open No.
No. 58) has excellent advantages as a carrier for an indirect agglutination reaction for detecting AIDS-related virus antibodies, for example, because it has few nonspecific reactions and can add properties depending on the application.

ゼラチン粒子へのHIV−p24及び/又はHIV−p41の感作
は、それ自体既知の方法を用いて行うことができ、例え
ばタンニン酸、グリタールアルデヒド、ビスジアジベン
ジジン、トリレンジイソシアネート、ジフロロジニトロ
ベンゼン、カルボジイミド類、キノン類、塩化クロム等
のいわゆるカップリング剤を使用して化学的に結合させ
る方法或いは物理的に吸着させる方法などによって行う
ことができる。
Sensitization of gelatin particles with HIV-p24 and / or HIV-p41 can be carried out using a method known per se, for example, tannic acid, glutaraldehyde, bisdiadibenzidine, tolylene diisocyanate, difluoro It can be performed by a method of chemically bonding or physically adsorbing using a so-called coupling agent such as dinitrobenzene, carbodiimides, quinones, and chromium chloride.

感作は弱アルカリ性の条件下で行えばよく、pH7.5〜
8.0の範囲内で行うのがよく、このpH範囲外で感作した
場合には一般に検出感度に優れた試薬は得られない。例
えば、後記実施例1又は2に記載の方法でゼラチン粒子
に各種pH条件下においてHIV−gp24又は参考としてHIV−
gp41を感作させた場合の感度を試験した結果を示せば下
記第1表のとおりである。
Sensitization may be performed under slightly alkaline conditions, and pH 7.5 to
The reaction is preferably performed within the range of 8.0. When sensitization is performed outside this pH range, a reagent excellent in detection sensitivity is generally not obtained. For example, HIV-gp24 or HIV-gp24 as a reference was added to gelatin particles under various pH conditions by the method described in Example 1 or 2 described below.
The results of testing the sensitivity when sensitizing gp41 are shown in Table 1 below.

第1表 感作pH 対照試験 力価試験 6.0 − − 7.0 − + 7.5 − + 8.0 − + 9.0 − + 10.0 + 測定不能 上記第1表において、−は凝集反応が生じなかったこ
とを、そして+は凝集反応が生じたことを示す。また、
対照試験とは、AIDS関連ウイルス抗体を全く含まない血
清希釈溶液を検体として用いた試験であり、この試験で
+の結果を示すのは偽反応である。一方、力価試験は抗
HIV−env(p41)タンパク抗体又は抗HIV−core(p24)
タンパク抗体を含む血清溶液を検体として用いた試験で
あり、十の数が多い程、の感度が高いことを示す。
Table 1 Sensitization pH control test Potency test 6.0 − − 7.0 − + 7.5 − + 8.0 − + 9.0 − + 10.0 + Unmeasurable In Table 1 above, -indicates that no agglutination reaction occurred, and + indicates Indicates that an agglutination reaction has occurred. Also,
The control test is a test using a serum diluted solution containing no AIDS-related virus antibody as a specimen, and a positive result in this test is a false reaction. On the other hand, the titer test
HIV-env (p41) protein antibody or anti-HIV-core (p24)
This is a test in which a serum solution containing a protein antibody is used as a specimen, and the larger the number, the higher the sensitivity.

以上述べた如くして得られるHIV−p24及び/又はHIV
−p41で感作したゼラチン粒子は通常、凍結乾燥した状
態で保存され、使用時に復元後、間接凝集反応に供せら
れる。
HIV-p24 and / or HIV obtained as described above
The gelatin particles sensitized with -p41 are usually stored in a lyophilized state, restored after use, and subjected to an indirect agglutination reaction.

本発明の感作ゼラチン粒子を用いる間接凝集反応によ
るAIDS関連ウイルス抗体の検出はそれ自体既知の間接凝
集反応法と全く同様にして行うことができ、例えばマイ
クロプレートのウエルに被検血清を入れてその希釈列を
作り、各ウエルに本発明の感作ゼラチン粒子を加えて混
合し、通常1〜2時間程度静置し、この感作ゼラチン粒
子の凝集像を観察して凝集の有無を判定することにより
行われる。
Detection of an AIDS-related virus antibody by indirect agglutination using the sensitized gelatin particles of the present invention can be performed in exactly the same manner as the indirect agglutination reaction known per se, for example, by placing a test serum in a well of a microplate. The dilution line is formed, and the sensitized gelatin particles of the present invention are added to each well and mixed, and the mixture is allowed to stand usually for about 1 to 2 hours. The aggregation image of the sensitized gelatin particles is observed to determine the presence or absence of aggregation. This is done by:

より具体的に述べれば、感作ゼラチン粒子浮遊液をAI
DS患者血清(検体)の二倍段階希釈液に96穴マイクロプ
レートのウエルに等量ずつ加え、混合し、室温に静置す
る。1〜3時間後、各穴を観察すると、希釈倍数の低い
穴には感作ゼラチン粒子が底一面に広がって、凝集像が
見られ、希釈倍数の高い所では、ボタン状となり凝集像
が認められない。一方、未感作ゼラチン粒子を用いた場
合には低希釈から高希釈の血清中いずれにも凝集像は見
られない。また、HIV抗体陰性ヒト血清について同様に
検査した場合には、感作ゼラチン粒子及び未感作ゼラチ
ン粒子のいずれを用いた場合にも凝集像は見られない。
More specifically, the sensitized gelatin particle suspension was
An equal volume of a two-fold serial dilution of DS patient serum (sample) is added to the wells of a 96-well microplate, mixed, and allowed to stand at room temperature. After 1 to 3 hours, when observing each hole, the sensitized gelatin particles spread all over the bottom in the hole with a low dilution factor, and an aggregated image was observed. I can't. On the other hand, when unsensitized gelatin particles were used, no aggregation image was observed in any of the low- to high-diluted serum. In addition, when the HIV antibody negative human serum was examined in the same manner, no aggregated image was observed when either the sensitized gelatin particles or the unsensitized gelatin particles were used.

これにより、検体中に、感作ゼラチン粒子上にHIV−p
24又はHIV−gp41あるいはp41と特異的に反応するHIV抗
体[抗HIV−core(p24)タンパク抗体、抗HIV−env(gp
41あるいはp41)タンパク抗体など]が存在することが
確認できる。
This allows HIV-p on the sensitized gelatin particles in the sample.
24 or an HIV antibody that specifically reacts with HIV-gp41 or p41 [anti-HIV-core (p24) protein antibody, anti-HIV-env (gp
41 or p41) protein antibody etc.] can be confirmed.

本発明の感作ゼラチン粒子は、AIDS関連ウイルス抗体
検出用試薬として、通常行なわれているように、未感作
ゼラチン粒子、標準血清(対照用陽性血清)、検体希釈
用溶液及び必要に応じて感作ゼラチン粒子の凍結乾燥品
溶解用溶液等と組合わせることによりキット化して実用
に供することができる。
The sensitized gelatin particles of the present invention may be used as a reagent for detecting an AIDS-related virus antibody, as usual, with unsensitized gelatin particles, standard serum (positive serum for control), a sample diluting solution, and A kit can be put into practical use by combining it with a solution for dissolving a lyophilized product of sensitized gelatin particles.

尚、本発明においてはp41をgp41に対して便宜上使用
したものであるが、正確にはgp41から糖の脱落したタン
パクを指すものである。
In the present invention, p41 is used for convenience with respect to gp41, but more accurately refers to a protein in which sugar is removed from gp41.

[実 施 例] 次に実施例により、本発明の感作ゼラチン粒子の調製
法及びそれを用いた間接凝集反応テストについてさらに
具体的に説明する。
[Examples] Next, the methods for preparing the sensitized gelatin particles of the present invention and the indirect agglutination test using the same will be described more specifically with reference to the following examples.

参考例 1 (1) HIV−env(gp41)タンパク抗原感作ゼラチン粒
子の調製 特開昭58−113754号公報の実施例1に記載された方法
に従って製造したゼラチン粒子を5ppmのタンニン酸を含
むpH7.2の0.15Mリン酸塩緩衝生理食塩液(以下0.15M P
BS(pH7.2)と略記する)中に粒子濃度が2.5%になるよ
うに分散させ、37℃で10分間加温した。この粒子を遠心
分離して回収し、生理食塩液で3回遠心分離法で洗浄し
てから0.15M PBS(pH8.0)に5%になるように分散さ
せた。
Reference Example 1 (1) Preparation of HIV-env (gp41) Protein Antigen Sensitized Gelatin Particles Gelatin particles produced according to the method described in Example 1 of JP-A-58-113754 were adjusted to pH 7 containing 5 ppm of tannic acid. .2 0.15M phosphate buffered saline (0.15MP
BS (abbreviated as pH 7.2)) so as to have a particle concentration of 2.5%, and heated at 37 ° C for 10 minutes. The particles were collected by centrifugation, washed three times with a physiological saline solution by a centrifugation method, and then dispersed in 0.15 M PBS (pH 8.0) so as to be 5%.

HIV−env(gp41)タンパク抗原を0.15M PBS(pH8.
0)で20μg/mlになるように希釈し、前記のタンニン酸
処理粒子分散液に等容のこの希釈液を混合して37℃で90
分間加温し、粒子にHIV−env(gp41)タンパク抗原を感
作させた。この感作ゼラチン粒子を生理食塩液で3回遠
心洗浄してから浮遊分散液(リン酸水素二ナトリウム・
12水塩1.920g、リン酸二水素カリウム0.291g、塩化ナト
リウム0.438g、アラビアゴム0.25g、グリシン1.5g、ア
ジ化ナトリウム0.1g及び正常家兎血清1.0mlを全量100ml
になるように、精製水に溶解した液)に分散させた。こ
の分散液を凍結乾燥し、HIV−env(gp41)タンパク抗原
感作ゼラチン粒子の凍結乾燥品を得た。
HIV-env (gp41) protein antigen was added to 0.15 M PBS (pH 8.
0), the mixture was diluted to 20 μg / ml with the above-mentioned tannic acid-treated particle dispersion, and an equal volume of this diluent was mixed.
After heating for 1 minute, the particles were sensitized with the HIV-env (gp41) protein antigen. The sensitized gelatin particles are centrifugally washed three times with a physiological saline solution, and then suspended in a suspension (disodium hydrogen phosphate.
1.920 g of dodecahydrate, 0.291 g of potassium dihydrogen phosphate, 0.438 g of sodium chloride, 0.25 g of gum arabic, 1.5 g of glycine, 0.1 g of sodium azide and 1.0 ml of normal rabbit serum in a total volume of 100 ml
(A solution dissolved in purified water). This dispersion was freeze-dried to obtain a freeze-dried product of HIV-env (gp41) protein antigen-sensitized gelatin particles.

(2) 被検血清希釈用溶液の調製 リン酸水素二ナトリウム・12水塩1.920g、リン酸二水
素カリウム0.291g、塩化ナトリウム0.438g、アラビアゴ
ム0.25g、アジ化ナトリウム0.15g、を全量100mlになる
ように精製水に溶解して血清希釈溶液とした。
(2) Preparation of test serum dilution solution 1.20 g of disodium hydrogen phosphate / 12 hydrate, 0.291 g of potassium dihydrogen phosphate, 0.438 g of sodium chloride, 0.25 g of gum arabic, 0.15 g of sodium azide To obtain a serum diluted solution.

(3) キットの調製 感作ゼラチン粒子の凍結乾燥品のほかに抗原未感作ゼ
ラチン粒子の凍結乾燥品及び対照用陽性血清の凍結乾燥
品も別途に調製し、感作ゼラチン粒子溶解用溶液及び被
験血清希釈用溶液を組合わせ、抗HIV−env(p41)タン
パク抗体検出用試験キットとした。
(3) Preparation of kit In addition to the freeze-dried product of the sensitized gelatin particles, a freeze-dried product of the antigen-unsensitized gelatin particles and a freeze-dried product of the control positive serum were separately prepared. The test serum dilution solution was combined to prepare a test kit for detecting anti-HIV-env (p41) protein antibody.

実施例 1 (1)HIV−core(p24)タンパク抗原感作ゼラチン粒子
の調製 実施例1で用いたHIV−env(gp41)タンパク抗原の代
わりにHIV−core(p24)タンパク抗原(セントコア社
製)を用い、同様な操作をくり返すことによりHIV−cor
e(p24)タンパク抗体抗原感作ゼラチン粒子を調製し
た。
Example 1 (1) Preparation of HIV-core (p24) Protein Antigen-Sensitized Gelatin Particles Instead of the HIV-env (gp41) protein antigen used in Example 1, HIV-core (p24) protein antigen (Centcore) And repeated the same operation to obtain HIV-cor
e (p24) protein antibody antigen-sensitized gelatin particles were prepared.

(2) キットの調製 実施例1と同様に試薬を用いHIV−core(p24)タンパ
ク抗体検出用試薬キットとした。
(2) Preparation of Kit An HIV-core (p24) protein antibody detection reagent kit was prepared using the same reagents as in Example 1.

実施例 2 実施例1及び2で得られた試験キットを用い、各種被
検血清について間接凝集反応テストを行なった。
Example 2 Using the test kits obtained in Examples 1 and 2, indirect agglutination tests were performed on various test sera.

得られた結果を下記第2表に示す。 The results obtained are shown in Table 2 below.

上記第2表に示す結果より、本発明に従うHIV−env
(gp41)タンパク抗原感作ゼラチン粒子及びHIV−core
(p24)タンパク抗原感作ゼラチン粒子を用いた間接凝
集反応は、抗HIV−env(gp41)タンパク抗体及び抗HIV
−core(p24)タンパク抗体の検出において、ウエスタ
ンブロット法と比べ同等の検出感度及び特異性を有する
ことがわかる。
From the results shown in Table 2 above, the HIV-env according to the present invention
(Gp41) Protein antigen-sensitized gelatin particles and HIV-core
(P24) Indirect agglutination using protein antigen-sensitized gelatin particles was performed using anti-HIV-env (gp41) protein antibody and anti-HIV
It can be seen that the detection of the -core (p24) protein antibody has the same detection sensitivity and specificity as the Western blot method.

[発明の効果] 本発明の試薬を用いれば、間接凝集反応により、多数
の検体を簡便な動作で迅速に測定することができる。例
えば、螢光抗体法あるいは酵素抗体法の場合、検査開始
から結果がでるまで洗浄も加え回数の操作を必要とする
が、本発明の試薬を用いた場合、希釈血清に加えるだけ
で、あとは結果がでるまで静置しておけばよく、非常に
簡便である。また、結果の判定には特殊な器材を何ら必
要とせず肉眼で判定することができる。これらの点は、
多数の検体の検査に適している。
[Effects of the Invention] By using the reagent of the present invention, a large number of samples can be quickly measured by a simple operation by an indirect agglutination reaction. For example, in the case of the fluorescent antibody method or the enzyme antibody method, washing and additional operations are required until the result is obtained from the start of the test, but when the reagent of the present invention is used, it is only necessary to add to the diluted serum, It is very convenient to leave it still until results are obtained. In addition, the result can be determined with the naked eye without any special equipment. These points are
Suitable for testing a large number of samples.

AIDS関連ウイルス抗体が検出された場合、AIDS関連ウ
イルスを体内に保有していると考えられるので、本発明
の試薬を用い、AIDS関連ウイルス抗体の有無を検査する
ことにより、接触及び輸血などによる感染を防止しうる
ので臨床上非常に有用である。
If an AIDS-related virus antibody is detected, it is considered that the virus has the AIDS-related virus in the body. Is very clinically useful because it can prevent

───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 昭64−57171(JP,A) 特開 昭63−128260(JP,A) 特開 昭62−182662(JP,A) 特開 昭62−278459(JP,A) ──────────────────────────────────────────────────続 き Continuation of front page (56) References JP-A-64-57171 (JP, A) JP-A-63-128260 (JP, A) JP-A-62-182662 (JP, A) JP-A-62-182662 278459 (JP, A)

Claims (1)

(57)【特許請求の範囲】(57) [Claims] 【請求項1】HIV核タンパクP24及び/又はHIV外殻タン
パクp41をpH7.5〜8.0において感作したゼラチン粒子か
らなることを特徴とする後天性免疫不全症候群関連ウイ
ルス抗体検出用間接凝集反応試薬。
1. An indirect agglutination reagent for detecting an acquired immunodeficiency syndrome-associated virus antibody, comprising gelatin particles sensitized with HIV nuclear protein P24 and / or HIV outer protein p41 at pH 7.5 to 8.0. .
JP62326774A 1987-12-25 1987-12-25 Virus antibody detection reagent Expired - Lifetime JP2587969B2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP62326774A JP2587969B2 (en) 1987-12-25 1987-12-25 Virus antibody detection reagent

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Application Number Priority Date Filing Date Title
JP62326774A JP2587969B2 (en) 1987-12-25 1987-12-25 Virus antibody detection reagent

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JPH01169360A JPH01169360A (en) 1989-07-04
JP2587969B2 true JP2587969B2 (en) 1997-03-05

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Country Link
JP (1) JP2587969B2 (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2512627B2 (en) * 1990-11-08 1996-07-03 株式会社トクヤマ Method for producing immunological agglutination reagent for long-term storage stability, and liquid for dissolving immunological agglutination particles used therefor

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4921787A (en) * 1987-05-01 1990-05-01 Cambridge Bioscience Corporation Detection of antibodies to human immunodeficiency virus by agglutination of antigen coated latex

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