流行性乙型脑炎多克隆抗体的制备及其囊膜E蛋白ELISA方法的建立  

Preparation of polyclonal antibody against Japanese encephalitis virus and the establishment of ELISA method using envelope E protein

作　　者：张宁[1,2] 金洪涛[1] 丁壮[2] 张瑞岩[1] 刘雯[2] 李勇[2] 连海[1] 夏志平[1] 

机构地区：[1]中国人民解放军军事医学科学院军事兽医研究所吉林省人兽共患病预防与控制重点实验室,吉林长春130062 [2]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《黑龙江畜牧兽医》2010年第10期10-14,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：中国农业部农业公益行为科研项目(200803015)

摘　　要：为了建立流行性乙型脑炎病毒(JEV)的检测方法,试验用纯化的乙型脑炎病毒免疫健康新西兰白兔制备多克隆抗体,并对多克隆抗体进行纯化和效价测定;以纯化原核表达的乙型脑炎重组结构蛋白(E蛋白)为包被抗原,通过对检测方法相关条件的优化,建立了检测乙型脑炎病毒抗体的间接ELISA方法。结果表明:该抗原不与猪其他疾病的阳性血清反应,具有良好的特异性;在批内重复试验中,变异系数小于10%。To establish a method for detecting the Japanese encephalitis virus（JEV）,New Zealand albino rabbits were immuned with purified JEV to obtain polyclonal antibody,which was purified and titrated.Through optimizing various factors affected the experiments,an indirect ELISA was established for the rapid detection of specific Japanese encephalitis virus（JEV） using a recombinant E protein as an antigen.The results showed that the E protein antigen had a good specificity,which did not take reaction with positive serum of other diseases of swine.In the duplicate tests,coefficient of variation was less than 10%.This research has provided a convenient method for the serum diagnosis of JEV.

关 键 词：乙型脑炎病毒 E蛋白 ELISA方法 检测 

分 类 号：S852.65[农业科学—基础兽医学]