猪圆环病毒2型间接ELISA抗体检测试剂盒的研制及初步应用

Development and Primary Application of Indirect ELISA Antibody Detection Kit for Porcine Circovirus Serotype 2

以大肠杆菌原核表达系统表达的猪圆环病毒2型Cap蛋白为抗原,建立猪圆环病毒2型间接ELISA检测方法,优化ELISA反应条件,研制猪圆环病毒2型ELISA抗体检测试剂盒。与商品化试剂盒相比,该检测试剂盒敏感性、特异性和符合率分别为95.12%、92.86%和94.55%;同时与猪圆环病毒1型(PCV1)、猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)等多种病毒阳性血清无交叉反应。试剂盒具有较好的重复性,在-20℃至少可保存1年以上,将其应用于临床血清样品的检测,结果安徽、广东、广西采样猪场的PCV2阳性率分别为100%、48.39%、100%。

Using the purified Cap recombinant protein as coating antigen of indirect ELISA,detecting kit was developed by the optimal reaction conditions of ELISA.In comparative experiments with commercial ELISA kit for detection of 55 serum samples,the sensitivity,specificity and coincidence rate of the established ELISA method were 95.12%,92.86% and 94.55% respectively.The established ELISA method had no cross-reactivity with the reference sera of other swine viruses（PCV1,CSFV,PRRSV,PPV,PRV）.The intra-assay variation coefficient was 2.3% to 4.5%,inter-assay variation coefficient was 1.12% to 2.63% respectively.The ELISA antibody detection kit could be stable at-20 ℃ at least for one year.66 serum samples of three pig farms from Anhui,Guangdong,Guangxi were detected by detection kit,and the positive rate was 100%,48.39%,100% respectively.