基于衣壳的竞争抑制酶联免疫吸附测定法的开发，用于检测牛和水牛血清中的牛免疫缺陷病毒抗体。

Development of a capsid based competitive inhibition enzyme-linked immunosorbent assay for detection of bovine immunodeficiency virus antibodies in cattle and buffalo serum.

作者信息

Bhatia S, Sood Richa, Bhatia A K, Pattnaik B, Pradhan H K

机构信息

High Security Animal Disease Laboratory, Indian Veterinary Research Institute, Anandnagar, Bhopal, Madhya Pradesh 462021, India.

出版信息

J Virol Methods. 2008 Mar;148(1-2):218-25. doi: 10.1016/j.jviromet.2007.11.008. Epub 2008 Jan 14.

DOI:10.1016/j.jviromet.2007.11.008

PMID:18192030

Abstract

The aim of this study was to develop a more specific and sensitive competitive inhibition ELISA (CI-ELISA) than the currently used indirect ELISA for detection of antibodies to bovine immunodeficiency virus (BIV) in cattle and buffaloes. Murine monoclonal antibodies (MAbs) were generated against a recombinant capsid (CA) protein of bovine immunodeficiency virus. Of the 13 anti-CA MAbs developed, MAb-9G10 was selected for CI-ELISA based on the maximum inhibition (98%) obtained with reference BIV antibody positive serum. Based on the distribution of percent inhibition of known negative sera (n=50), a cut-off value was set at 40% inhibition. The MAb-based CI-ELISA showed much higher agreement (concordance: 95.4%) than the indirect ELISA (concordance: 77.8%) with Western blot. Out of 672 sera of cattle and buffaloes tested by CI-ELISA from four states of India, 22% (113/516) of cattle and 19% (30/156) of buffalo were sero-positive for BIV with an overall seroprevalence of 21% (143/672) in India.

摘要

本研究的目的是开发一种比目前用于检测牛和水牛体内牛免疫缺陷病毒（BIV）抗体的间接ELISA更特异、更灵敏的竞争抑制ELISA（CI-ELISA）。针对牛免疫缺陷病毒的重组衣壳（CA）蛋白产生了鼠单克隆抗体（MAb）。在开发的13种抗CA MAb中，基于参考BIV抗体阳性血清获得的最大抑制率（98%），选择MAb-9G10用于CI-ELISA。根据已知阴性血清（n=50）抑制率百分比的分布，将临界值设定为40%抑制率。基于MAb的CI-ELISA与蛋白质印迹法的一致性（符合率：95.4%）远高于间接ELISA（符合率：77.8%）。在印度四个邦用CI-ELISA检测的672份牛和水牛血清中，22%（113/516）的牛和19%（30/156）的水牛BIV血清学呈阳性，印度的总体血清阳性率为21%（143/672）。