猪繁殖与呼吸综合征病毒Nsp7间接ELISA检测方法的建立

Establishment of indirect ELISA for detecting porcine reproductive and respiratory syndrome virus with Nsp7 as coating antigen

为建立猪繁殖与呼吸综合征病毒(PRRSV)抗体的检测方法,本研究以重组Nsp7作为包被抗原,通过优化反应条件,建立PRRSV抗体间接ELISA检测方法。该检测方法与猪瘟等常见的6种猪病毒病的阳性血清无交叉反应,显示出良好的特异性。其检测标准血清的最低稀释倍数为1∶3 200,显示出良好的敏感性。批内重复性试验的变异系数小于5%,批间重复性试验的变异系数小于10%,显示出良好的重复性。采用该检测方法检测400份临床血清样品,实验结果表明该方法与商品化的IDEXX PRRSV抗体检测试剂盒符合率达到95.0%～96.8%;与LSI PRRSV抗体检测试剂盒符合率达到94.0%～96.5%。

To establish the method for detection of the antibody against porcine reproductive and respiratory syndrome virus （PRRSV）, the indirect ELSA was set up with the Nsp7 recombinant protein as coating antigen. Under the optimization conditions, this method was no cross-reaction with other 6 porcine viruses and the limit detection was 1：3,200 for the standard positive PRRSV serum. The reproducibility tests showed that the coefficients of variation for intra- and inter-assay were lower than 5% and 10%, respectively. Tested on 400 clinical serum samples, the results displayed that the coincidence rates of this method were 95.0% to 96.8% with the IDEXX commercial PRRSV antibody detection kit and 94.0% to 96.5% with the LSI PRRSV antibody detection kit, respectively.