猪乙脑病毒CSF.XZ-2D株的分离鉴定及其体外培养特性研究  

Isolation and propagation of a field Japanese encephalitis virus isolate CSF.XZ-2D from swine

作　　者：滕蔓[1] 罗俊[1] 樊剑鸣[2] 王兴涛[1] 王方雨[1] 柴书军[1] 杨继飞[1] 张改平[1] 

机构地区：[1]河南省农业科学院农业部动物免疫学重点开放实验室河南省动物免疫学重点实验室,河南郑州450002 [2].郑州大学公共卫生学院,河南郑州450001

出　　处：《中国预防兽医学报》2012年第4期274-278,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家公益性行业(农业)科研专项经费(200803015、201203082)

摘　　要：为了解河南省规模化猪场日本乙型脑炎病毒(JEV)流行株的特征,本研究对临床流产胎儿脑脊液进行病毒盲传分离,经RT-PCR等分子生物学方法及乳鼠感染试验,分离鉴定了1株基因Ⅲ型JEV株,命名为CSF.XZ-2D。采用BHK-21细胞对该分离株的体外培养特性进行研究,结果表明,在25 cm2细胞瓶中,用1.25×106个细胞/瓶的初始量制备细胞单层,以较低剂量的病毒液吸附细胞并洗涤弃去游离病毒粒子,可在感染后60 h达到最大增殖量,病毒效价为105.0TCID50/0.1 mL以上。本研究为其他JEV分离株的体外培养提供了参考。To investigate the features of Japanese encephalitis virus（JEV） currently prevalent in pig farm in Henan province,the cerebrospinal fluid（CSF） samples collected from aborted fetuses were blind passaged on BHK-21 cells.A JEV was isolated and characterized by RT-PCR,immunofluorescence assay and in animal tests.The isolate was named as CSF.XZ-2D and the phylogenetic analysis showed that it belongs to genotype III.The conditions for the culture of JEV CSF.XZ-2D were optimized in BHK-21 cells.The virus replication curves showed that the highest virus titer achieved above 105.0 TCID50/0.1 mL at 60 hours post-inoculation,which indicated that our results would facilitate improvements for the in vitro culture of other JEV strains.

关 键 词：日本乙型脑炎病毒 分离鉴定 脑脊液 BHK-21细胞 体外培养 

分 类 号：S852.65[农业科学—基础兽医学]