一株流产猪胎来源的GI JEV的分离、鉴定与分子特征  

Isolation,Identification and Molecular Characterization of A Japanese Encephalitis Virus Genotype Ⅰ from Aborted Pig Fetuses

作　　者：郭爽 王欣[1,2] 庞琳琳[1] 张俊杰 李宗杰 李蓓蓓 刘珂[1] 邵东华[1] 邱亚峰[1] 马志永[1] 魏建超[1] GUO Shuang;WANG Xin;PANG Linlin;ZHANG Junjie;LI Zongjie;LI beibei;LIU Ke;SHAO Donghua;QIU Yafeng;MA Zhiyong;WEI Jianchao(Shanghai Veterinary Research Institute,CAAS,Shanghai 100141,China;Yangtze University,Jingzhou 434000,China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]长江大学,荆州434000

出　　处：《中国动物传染病学报》2022年第4期110-116,共7页Chinese Journal of Animal Infectious Diseases

基　　金：上海市科技兴农项目(X2021-02-08-00-12-F00770,X2022-02-08-00-12-F01195)。

摘　　要：2015年从上海市奉贤区某猪场采集疑似感染乙脑的猪病样品,通过RT-PCR方法对病料进行检测,检测阳性样本采用BHK-21细胞进行病毒分离,利用间接免疫荧光(IFA)技术对分离株进行鉴定。结果显示,从流产胎儿的脑组织中成功分离到一株JEV,命名为SD12。SD12株感染BHK-21细胞,48 h均发生病变,病毒滴度为1.25×10^(6) PFU。分别采用脑内(IC)和腹腔内(IP)感染小鼠测得脑内感染半数致死量(LD_(50))为2.5 PFU,经腹腔感染的LD_(50)为10^(2.7) PFU,表明该分离毒株SD12具有强的神经毒力和神经侵袭力。对SD12株的全基因进行克隆测序和进化分析,结果显示,该分离株基因全长10965 bp,属于基因I型JEV,与基因Ⅲ型经典弱毒疫苗株SA14-14-2株基因核苷酸同源性为88.6%,氨基酸同源性为97.4%,其中E基因同源性87.5%,氨基酸同源性为97.2%。本研究为猪乙型脑炎的防控与疫苗研发奠定了基础。In 2015,Brain samples were collected from aborted fetuses suspicious of Japanese encephalitis virus(JEV)in Fengxian area,Shanghai,and examined by RT-PCR.The PCR positive samples were inoculated into BHK-21 cells for virus isolation and the presenc3 of the JEV was confi rmed with IFA.As a result,a JEV isolate was obtained and designated as SD12 strain.The SD12 strain propagated in BHK-21 cells and infected cells formed CPE at 48 h post infection with the titer of 1.25×10^(6) PFU.Inoculation of the SD12 strain into mice caused deaths and the median lethal doses(LD_(50))were 2.5 PFU for intracerebral injection(IC)and 10^(2.7) PFU for peritoneal injection(IP),which indicated that the SD12 strain had a strong virulence and nerve invasiveness.The entire gene of the SD12 strain was cloned and sequenced for phylogenetic analysis.The results showed that the full length of the SD12 strain genome was 10965 bp and belonged to the genotype I(GI).Moreover,the genomic sequence of the SD12 strain was also compared with that of the GⅢclassic attenuated vaccine strain SA14-14-2.The SD12 strain shared 88.6%nucleotide homology and 97.4%amino acid homology,of which E gene had 87.5%nucleotide homology and 97.2%amino acid homology.In conclusion,the results of the study laid the foundation for further study on vaccine development and prevention of swine JEV.

关 键 词：乙型脑炎病毒、基因Ⅰ型 分离鉴定 进化分析 

分 类 号：S852.65[农业科学—基础兽医学]