狂犬病病毒中和抗体ELISA技术的建立

将狂犬病病毒(RV)糖蛋白(G蛋白)中和抗原表位串联表达的重组蛋白作为抗原,建立了检测RV中和抗体的间接ELISA技术。结果表明,最佳抗原包被量为2μg/孔,被检血清最佳稀释倍数为1:200。该方法与快速荧光灶抑制试验(RFFIT)的阳性符合率为88%,阴性符合率为96%。特异性试验表明,该抗原不与犬腺病毒I型、犬细小病毒、犬瘟热病毒、犬副流感病毒和犬冠状病毒阳性血清发生交叉反应,具有良好的特异性。板内和板间重复性试验的平均变异系数分别为2.7%和4.2%,具有良好的重复性,为动物RV中和抗体检测提供了简单快捷的检测方法。

Indirect ELISA for Determination of Neutralizing Antibody of Rabies Virus

An indirect ELISA assay for determination of neutralizing antibody of rabies virus was established by using the recombinant G protein as antigen.The recombinant G protein was obtained by expressing the spliced gene fragments that encode polypeptides comprising the linear neutralization sites of the G protein.The results showed that the amount of recombinant G protein for coating was 2 μg per well and the dilution for the detecting sample was 1:200.The positive coincidence rate and negative coincidence rate of samples between the established ELISA and rapid fluorescent focus inhibition test(RFFIT) was 88% and 96% respectively.The ELISA did not cross-react with sera positive for canine adenovirus type I,canine parvovirus,canine distemper virus,canine parainfluenza virus and canine coronavirus,showing good specificity.The intra-assay and inter-assay coefficient of standard variation was 2.7% and 4.2% respectively.The indirect ELISA would be useful for detecting neutralizing antibody of rabies virus in animals.