应用间接ELISA检测鸡痘病毒抗体方法的建立

Development of indirect ELISA for detection of antibodies against fowlpox virus

应用鸡胚成纤维细胞(CEF)扩增鸡痘病毒(Fowlpox virus,FPV)并提纯后作为抗原建立了具有较高特异性和灵敏性的间接ELISA方法.通过方阵滴定法来确定抗原最佳包被浓度为2.7μg/孔,待检血清最佳稀释倍数为1:100,其阳性临界值为OD≥0.113.将400份FPV免疫实验鸡血清用本方法进行检测,其阳性检出率为81.25%(325/400).此外,将该方法与琼脂扩散试验进行比较检测血清样品,结果显示本方法的灵敏度比琼脂扩散试验灵敏400～800倍,而且还有特异性强,操作简便、快速等优点.

The FPV propagated on CEF was harvested as antigen to develop an indirect ELISA for detecting antibodies of fowlpox virus. The optimal coating concentration of antigen was 2.7ug of FPV protein per well, the serum sample were diluted to 1 ： 100 and the cutoff was determined to be 0. 113. The indirect ELISA detected a total of 400 serum sample originated from chickens immunized with FPV or rFPV, and the positive efficacy was 81.25 % （325/400）. The test also revealed that indirect ELISA was 400 to 800 times more sensitive than AGP. Otherwise, it was simple, convenient, specific and rapid for detecting antibodies of fowlpox virus.