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.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
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)
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
Japanese encephalitis virus
viral isolation and identification
cerebrospinal fluid
BHK-21 cell
in vitro culture
