猪瘟病毒重组E2蛋白PPA-ELISA抗体检测试剂盒的研制及应用

Development and application of PPA-ELISA for detecting antibodies to classical swine fever virus using recombinant E2 antigen

以纯化的猪瘟病毒(CSFV)重组E2蛋白为包被抗原,辣根过氧化物酶标记葡萄球菌A蛋白为二抗,组装了检测CSFV抗体的PPA-ELISA诊断试剂盒。该试剂盒与其他7种常见猪病毒(PRRSV、PPV、JEV、PCV-2、PEDV、PRV、TGEV)及牛病毒性腹泻病毒(BVDV)的阳性血清不发生交叉反应;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%;与间接血凝试验相比较,符合率、敏感性和特异性分别为86.2%,85.4%,87.2%;与IDEXX ELISA试剂盒相比较,符合率、敏感性和特异性分别为91.3%,93.2%,88.7%。表明本试验研制的E2-PPA-ELISA抗体检测试剂盒具有良好的重复性、敏感性和特异性,将为CSFV的快速诊断、免疫猪群抗体水平监测和CSFV流行病学调查提供一种快速、简便的血清学诊断方法。

To develop a method to detect the antibody of classical swine fever virus（CSFV）,an indirect ELISA kit was developed using the purified recombinant E2 protein as coating antigen and HRP-SPA as the second antibody.The E2-PPA-ELISA kit showed no cross-reaction with the positive sera of other eight virus（PRRSV,PPV,JEV,PCV-2,PEDV,PRV,TGEV,BVDV）.Coefficient of variability percent（C.V %） of intro-batch and inter-batch duplicative tests was less than 5% and 10%,respectively.Comparing with the indirect hemagglutination test and IDEXX ELISA kit,the concordance was 86.2% and 91.3%,the sensitivity was 85.4% and 93.2%,the specificity was 87.2 % and 88.7%.Therefore,the E2-PPA-ELISA kit has good specificity,sensitivity and repeatability,could be used as a simple and rapid serology detection method for monitoring of anti-CSFV antibody and epidemiologic survey of CSFV.