狂犬病病毒中和抗体ELISA技术的建立  

Indirect ELISA for Determination of Neutralizing Antibody of Rabies Virus

作　　者：罗金燕[1] 王水明[2] 李向东[1] 沈阳[1] 邱亚峰[1] 史子学[1] 丁铲[1] 艾峰[2] 刘学辉[2] 张帆[2] 刘佩红 马志永[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]南京出入境检验检疫局,江苏南京210001 [3]上海动物疫病预防控制中心,上海201103

出　　处：《中国动物检疫》2011年第1期48-51,共4页China Animal Health Inspection

基　　金：江苏出入境检验检疫局科研项目(2007KJ02);上海市科技兴农重点攻关项目(沪农科攻字(2006)第5-4号)

摘　　要：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。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.

关 键 词：狂犬病病毒 G蛋白 中和抗体 间接ELISA 

分 类 号：S852.659.5[农业科学—基础兽医学]