猪伪狂犬病病毒抗体间接ELISA检测方法的建立

Establishment of indirect ELISA for detection of antibodies against pseudorabies virus

为建立一种快速检测猪伪狂犬病病毒(PRV)的血清学方法,本实验采用原核重组表达的g E蛋白作为包被抗原,并对各反应条件进行优化,建立了检测PRV血清抗体的间接ELISA方法。以该方法检测PRV阳性血清和其它猪病病原阳性血清,除了PRV阳性血清呈阳性外,对其它猪病病原阳性血清的检测结果均为阴性,表明该方法具有良好的特异性。批内和批间重复试验的最大变异系数分别是5.95%和9.62%。此外,使用该方法对250份临床血清进行检测,与国内外4种商品化试剂盒的检测结果比较符合率最高为83.6%。表明该方法可以用于PRV的临床检测,为PRV流行病学调查和防控提供了一种快速有效的检测方法。

In this study, an indirect-ELISA （gE-ELISA） for the detection of the antibody against pseudorabies virus （PRV） in serum was established using the prokaryotic expressed glycoprotein E （gE） as coating antigen. The gE-ELISA was high specificity for detection of the PRV antibody, but no cross-reactions to the positive sera of classical swine fever virus, transmissible gastroenteritis virus, porcine reproductive and respiratory syndrome virus, porcine circovirus type 2 virus, porcine epidemic diarrhea virus, Actinobacilluspleuropneumoniae, Streptococcus suis type 2. The coefficient of variation of intra-assay and inter batch-assay was lower than 5.95% and 9.62%, respectively. Moreover, comparison of the established gE-ELISA with four commercial kits for detection of 250 clinical samples to PRV, the results showed that the highest concordance rate was 83.6%. It would provide a rapid and effective assay for detection of PRV infection and serosurvey.