为建立一种检测成本低、操作简便的蓝舌病血清抗体监测方法,本研究通过制备群特异性的蓝舌病多克隆检测抗体和优化抗原固定技术,建立了蓝舌病多克隆抗体C-ELISA检测方法.通过对150份牛羊已知阴性血清的检测确定该检测方法的cut-off值为40%;对58份已知阳性血清样品同时进行中和实验和C-ELISA检测,检测结果经SPSS软件进行t检测统计分析,Sig.(双侧)=0.651,相关性0.912,相关性显著.对24个BTV血清型阳性血清,2份牛BVD阳性血清,2份羊口蹄疫阳性血清,2份羊痘阳性血清进行检测.结果表明,24个BTV血清型阳性血清为阳性,其他血清全部为阴性.对150份已知阴性血清和55份已知阳性血清用中和试验、美国VMRD公司试剂盒和本方法分别进行检测.结果表明,本检测方法与中和试验检测结果符合率达100%,进口试剂盒符合率为96.7%.
In order to establish a low cost and simple operation method in this research through prepa- ration of specificity of bluetongue polyclonal detection antibody and antigen fixed technology, blue- tongue polyclonal antibody C-ELISA detection method was established. By detecting 150 cows and sheep negative serum, the cut-off value was 40%. Fifty-eight positive serum samples were detected by VNT and C-ELISA at the same time, and statistically analyzed by SPSS software. The results showed that Sig. (bilateral) = 0.651, correlation was 0. 912; 24 type positive serum of BTV, 2 BVD positive serum of cow, 2 FMDV positive serum of sheep and 2 positive serum of goat-pox were all detected by C-ELISA. The results showed that the 24 type BTV positive serum were all positive and other serum all negative. One hundred and fifty known negative serum and fifty-five known positive serum were tested by VNT, American VMRD company kit and this method. The results showed that this method was completely consistent with the result of VNT.