猪繁殖与呼吸综合征病毒抗体的荧光微球免疫学检测方法的建立  

Development of a fluorescent microbead-based immunoassay for the detection of porcine reproductive and respiratory syndrome virus IgG

作　　者：曾梦[1,2] 纪方晓 冀池海 陈万里 张桂红[1,2] 王衡 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]广东省兽医临床重大疾病综合防控重点实验室,广东广州510642

出　　处：《中国兽医科学》2017年第12期1497-1502,共6页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2016YFD0500707);广东省高等学校优秀青年教师培养计划项目(YQ2015030)

摘　　要：为了建立一种高通量、重复性好、特异性高的猪繁殖与呼吸综合征病毒(PRRSV)抗体的快速检测方法,本研究用纯化后的PRRSV非结构蛋白7(Nsp7)重组蛋白作为偶联抗原建立PRRSV抗体的荧光微球免疫学检测方法。对检测方法进行分析结果表明,批内、批间变异系数分别为3.2%、4.2%,与其他常见猪病阳性血清无交叉反应。当单抗浓度低至1 ng/mL时,该方法仍具有高灵敏度。本研究建立的荧光微球免疫学检测方法能快速检测临床样本PRRSV抗体,为PRRSV检测提供了一种新方法。In order to establish a high-throughput,reproducible and highly specific method to detect swine reproductive and respiratory syndrome virus（PRRSV） antibodies,the purified non-structural protein 7 （Nsp7） recombinant protein was used as an antigen to establish a fluorescent microsphere-based immunoassay（FMIA）.SDS-PAGE and Western-blot analysis showed that PRRSV Nsp7 recombinant was soluble and had good antigenicity. The intra-assay and inter-assay coefficient of variation ofMFIA was 3.2% and 4.2%,respectively,and there were no cross-reactions with the positive sera of other swine diseases. When the concentration of mAb was as low as 1 ng/mL,the method still had high sensitivity. These results indicate that the FMIA is rapid,sensitive and specific for detecting antibodies against swine PRRSV.

关 键 词：猪繁殖与呼吸综合征病毒 非结构蛋白7 原核表达 荧光微球免疫学检测方法 

分 类 号：S852.659.6[农业科学—基础兽医学]