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用单克隆抗体阻断酶联免疫吸附试验检测 PMV-1 特异性抗体。

Detection of PMV-1 specific antibodies with a monoclonal antibody blocking enzyme-linked immunosorbent assay.

机构信息

Division of Poultry, The National Veterinary Institute, Uppsala, Sweden.

出版信息

Avian Pathol. 1996 Dec;25(4):691-703. doi: 10.1080/03079459608419175.

DOI:10.1080/03079459608419175
PMID:18645892
Abstract

A highly reproducible monoclonal antibody (Mab) blocking ELISA (B-ELISA) has been developed and evaluated for the detection of NDV-specific antibodies. The Mab utilised is specific for a conserved PMV-1 serotype-specific epitope, as demonstrated by the indirect immunoperoxidase test. It reacted with all strains representing different serogroups within the PMV-1 serotype, but not with any strain belonging to other PMV serotypes. Sensitivity and specificity of the B-ELISA were compared with the haemagglutina-tion inhibition test (HI). Blocking and HI antibodies were detected in sera of chickens 8 days post-experimental infection. The B-ELISA proved consistently more sensitive than the HI test. In another survey, 62 sera from experimentally vaccinated chickens were tested; 95.2% proved positive by B-ELISA, 85.5% by indirect ELISA and 74% by HI test. When 504 field sera from vaccinated chickens and turkeys were tested, 98% were positive by B-ELISA, and 69% by HI. The specificity was evaluated by testing 1066 samples from NDV-free flocks, all of which proved negative by both methods. Other advantages of the B-ELISA include easy standardization and quality control, and ability to test sera from any species (including exotic or wild birds as well as mammals). The use of low dilution serum or egg-yolk samples makes the test quick and easy to perform and suitable for large-scale screening.

摘要

已开发并评估了一种高度可重现的单克隆抗体(Mab)阻断 ELISA(B-ELISA),用于检测 NDV 特异性抗体。所使用的 Mab 针对 PMV-1 血清型特有的保守表位具有特异性,这一点通过间接免疫过氧化物酶试验得到了证明。它与代表 PMV-1 血清型内不同血清群的所有毒株反应,但与属于其他 PMV 血清型的任何毒株均不反应。B-ELISA 的敏感性和特异性与血凝抑制试验(HI)进行了比较。在实验感染后 8 天的鸡血清中检测到阻断和 HI 抗体。B-ELISA 被证明始终比 HI 试验更敏感。在另一项调查中,对 62 份来自实验接种鸡的血清进行了测试;B-ELISA 阳性率为 95.2%,间接 ELISA 阳性率为 85.5%,HI 试验阳性率为 74%。当对来自接种鸡和火鸡的 504 份现场血清进行测试时,B-ELISA 阳性率为 98%,HI 阳性率为 69%。通过测试来自无 NDV 鸡群的 1066 个样本评估了特异性,两种方法均证明这些样本均为阴性。B-ELISA 的其他优点包括易于标准化和质量控制,以及能够测试来自任何物种(包括外来或野生鸟类以及哺乳动物)的血清。使用低稀释血清或卵黄样本可使测试快速简便,适合大规模筛查。

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