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用于抗原检测的捕获ELISA中,单克隆和多克隆抗呼吸道合胞病毒(RS)抗体的选择与性能。

The selection and performance of monoclonal and polyclonal anti-respiratory syncytial virus (RS) antibodies in capture ELISAs for antigen detection.

作者信息

Clayton A L, Albert Z I, Chantler S M

机构信息

Wellcome Research Laboratories, Beckenham, Kent, U.K.

出版信息

J Virol Methods. 1987 Sep;17(3-4):247-61. doi: 10.1016/0166-0934(87)90135-2.

Abstract

Six monoclonal antibodies directed against fusion protein (F) or nucleoprotein (NP) of respiratory syncytial virus (RS) have been investigated in an antigen capture ELISA for virus detection. The potency, spectrum and pattern of reactivity were investigated with the intention of selecting antibodies reacting with RS-common antigen determinants and with complementary rather than competitive activity. Two anti-F protein antibodies satisfied these criteria and were used with enzyme amplified detection in a two site monoclonal assay (MCA/MCA) or as detectors with a polyclonal antibody as capture (PCA/MCA). Comparative studies were performed with immunofluorescence (FA) as the reference test and nasopharyngeal aspirates processed in different ways. The PCA/MCA assay was superior to that using monoclonal antibodies alone and gave results comparable to the reference method. However, the apparent sensitivity related to FA varied with the type of sample processing used. These results emphasise the need for a critical analysis of the factors which can influence the sensitivity of a particular assay system before judgements on relative sensitivity are made.

摘要

在一种用于病毒检测的抗原捕获酶联免疫吸附测定(ELISA)中,对六种针对呼吸道合胞病毒(RS)融合蛋白(F)或核蛋白(NP)的单克隆抗体进行了研究。研究了其效价、反应谱和反应模式,目的是选择与RS共同抗原决定簇反应且具有互补而非竞争活性的抗体。两种抗F蛋白抗体符合这些标准,并在两点单克隆测定(MCA/MCA)中用于酶放大检测,或作为检测器与多克隆抗体作为捕获剂(PCA/MCA)一起使用。以免疫荧光法(FA)作为参考试验,并对以不同方式处理的鼻咽抽吸物进行了比较研究。PCA/MCA测定法优于单独使用单克隆抗体的方法,其结果与参考方法相当。然而,与FA相关的表观灵敏度随所用样品处理类型而变化。这些结果强调,在对相对灵敏度做出判断之前,需要对可能影响特定测定系统灵敏度的因素进行批判性分析。

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