中华按蚊源日本脑炎病毒的分离鉴定及其分子特征的分析

发布时间：2018-03-28 11:03

  本文选题：日本脑炎病毒　切入点：分离鉴定　出处：《中国兽医科学》2017年01期

【摘要】：为了解上海市蚊源日本脑炎病毒(JEV)的分子特征,2014年从上海市嘉定区猪场采集蚊媒样品1万余份,采用BHK21和C6/36细胞培养法进行病毒分离,通过RT-PCR和IFA技术对分离株进行鉴定。结果表明,分离到1株JEV,命名为JD-15,源自中华按蚊。用JD-15株分别感染BHK21和C6/36细胞,72 h后所有细胞均发生明显病变(CPE),其细胞半数感染量(TCID50)为1.96×106PFU。用分离株JD-15脑内接种3周龄乳鼠,结果测得半数致死量(LD50)为4.2×101PFU,表明该毒株具有较强的神经毒力。对JD-15株的E基因进行克隆测序及进化分析,结果显示,该株JEV属于基因Ⅲ型,与减毒活疫苗株SA-14-14-2的E基因核苷酸序列的同源性为98.1%,氨基酸序列的同源性为97.5%。本研究为乙型脑炎的流行病学调查提供了科学依据,对上海市蚊虫及蚊媒病的防治具有重要意义。
[Abstract]:In order to understand the molecular characteristics of Japanese encephalitis virus (JEV) from Shanghai mosquitoes, more than 10 000 mosquito vector samples were collected from Jiading District Pig Farm in 2014. The virus was isolated by BHK21 and C6 / 36 cell culture. The isolates were identified by RT-PCR and IFA. A strain of JEV, named JD-15, was isolated from Anopheles sinensis. BHK21 and C6 / 36 cells were infected with JD-15 strain for 72 h, respectively. Results LD50 was 4.2 脳 10 ~ (-1) PFU, which indicated that the strain had strong neurotoxicity. The E gene of JD-15 strain was cloned and sequenced. The result showed that the strain JEV belonged to type 鈪，

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