BHK-21单细胞克隆敏感株筛选及日本脑炎病毒分离  

Screening of BHK-21 Monoclonal Cell Sensitive Strain and Isolation of Japanese Encephalitis Virus

作　　者：向卫军[1] 杨涛涛[1] 李润成[1] 余兴龙[1] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128

出　　处：《动物医学进展》2017年第5期22-25,共4页Progress In Veterinary Medicine

基　　金：湖南省科技厅重点课题(2012NK2001)

摘　　要：为筛选出日本脑炎病毒(JEV)敏感度高的BHK-21细胞株,采用终点稀释法从母代BHK-21混合细胞中分离单细胞克隆株,应用接种BHK-21克隆株方法,从JEV阳性蚊子样品研磨液中分离病毒,筛选出的3号单细胞克隆株经3次传代后细胞产生明显病变,分离到1株JEV,5号单克隆株培养物核酸检测呈阳性,但是不产生细胞病变,母代混合细胞没有病变,JEV核酸检测阴性。根据1、3型JEV的prM与E基因设计4对引物初步鉴定分离株基因型,鉴定分离株为1型JEV。综合以上结果,构建的BHK-21单细胞克隆株比BHK-21混合细胞对分离JEV株敏感,更适合JEV的分离与培养。To screen single clone cell line for cultivation of JEV with high sensitivity, dilution method was used to isolate single clone line from BHK-21 heterogeneous cells. The obtainted single cell clones were subjected to JEV positive mosquito sample.The screened No.3 monoclonal cell line, 1 strain of JEV was obtained after 3 passages.The No.5 monoclonal strain cultured in nucleic acid testing was positive,but didn＇t produce cytopathic effect.The heterogeneous cells showed no CPE and nucleic acid test was negative.According to the type 1,3 JEV prM and E genes, four pairs of primers were designed to identify the isolated strains,and the isolates were identified as type 1 JEV.These results suggested the BHK-21 monoclonal cell line is more suitable to cultivate JEV compared to BHK-21 heterogeneous cells.

关 键 词：BHK_21单克隆细胞株 病毒分离鉴定 1、3型JEV分型引物 

分 类 号：S852.65[农业科学—基础兽医学]