浙江省分离到基因Ⅲ型乙型脑炎病毒  

Identification of genotype Ⅲ Japanese encephalitis virus isolated in Zhejiang province

作　　者：严菊英[1] 王建跃[2] 王忠发[2] 喻忠文 俞仲卿 周佳悦[1] 张严峻[1] 

机构地区：[1]浙江省疾病预防控制中心病毒所,杭州310051 [2]舟山市疾病预防控制中心传染病防制所 [3]定海区疾病预防控制中心微生物所 [4]义乌市疾病预防控制中心传染病防制所

出　　处：《中华预防医学杂志》2012年第8期722-727,共6页Chinese Journal of Preventive Medicine

摘　　要：目的 分析1982—1983年浙江省媒介蚊虫中流行性乙型脑炎病毒（JEV）的分子特征与基因型别。方法 于1982—1983年从浙江省定海区和义乌市采集蚊虫样本3188只，用于病毒分离。应用C6/36细胞分离病毒，血凝抑制试验鉴定分离株。于2011年对JEV分离株活化，应用空斑形成单位（PFU）法测定病毒滴度；进行乳鼠脑内接种实验；对病毒PrM和E基因区扩增、测序，用生物信息软件将分离株与JEV疫苗株SA14-14-2和浙江2007—2010年JEV进行序列分析比较、并构建进化树。结果 从捕获的3188只蚊虫中，分离出11株可以引起C6/36细胞规律病变的病毒，病变时间为72 h，病毒滴度为2.5～6.47 lg PFU/ml，乳鼠脑内接种病毒后72 h死亡。用病毒PrM区和E基因进行的基因分型分析，浙江1982—1983年分离株属于JEV基因Ⅲ型, 而浙江2007—2010年分离株则属于JEV基因I型。对5株分离株E基因进行分析，病毒间核苷酸同源性在98.9%以上，氨基酸同源性在99.8%以上，5株病毒与疫苗株SA14-14-2之间核苷酸同源性在97.7%以上，氨基酸同源性为99.2%，共有10个共同的氨基酸差异位点，与2007—2010年浙江省JEV相比，核苷酸同源性为87.7%～87.9%，氨基酸同源性在98.8%以上。结论 1982—1983年从浙江省定海和义乌蚊虫中分离到的JEV属于基因Ⅲ型毒株，为浙江省首次报道。Objective To investigate the molecular characteristics and genotype of Japanese encephalitis virus （JEV） found in vector mosquitoes in Zhejiang province from 1982 to 1983. Methods A total of 3188 mosquitoes were collected in Dinghai district and Yiwu city in Zhejiang province, during year 1982 and 1983. The virus was isolated by C6/36 cell, and then identified by hemagglutination inhibition test. The isolated strains were activated in year 2011, and plaque forming unit （PFU） were applied to test the virus titer. The suckling rats were tested under intracranial inoculation, where PrM and E genes were amplified and sequenced. Their nucleotide and amino acid sequences were analyzed and compared with the JEV vaccine strain SA14-14-2 and the JEV isolated in Zhejiang province during 2007 and 2010; and phylogenetic tree were constructed by bioinformatic software. Results From the 3188 mosquitoes captured, eleven virus strains were isolated and found to be able to cause cytopathogenic effect （CPE） in C6/36 cells within 72 hours. Virus titer ranged from 2.5 to 6.47 lg PFU/ml. The suckling rats would die within 72 hours since the inoculation. The phylogenetic analysis with the PrM and E genes showed that the JEV isolated in Zhejiang during 1982-1983 belonged to genotype Ⅲ; while the JEV isolated in Zhejiang during 2007-2010 belonged to genotype Ⅰ. The analysis of E genes from 5 isolated strains found that the homology of nucleotide sequence was over 98.9%, and the homology of amino acid sequence was over 99.8%. The compare between the 5 virus strains and the vaccine strain SA14-14-2 found 10 common amino acid variation sites, and showed that the homology of nucleotide sequence was over 97.7%, and the homology of amino acid sequence was 99.2%. The compare between the 5 virus strains and the JEV isolated in Zhejiang during 2007 and 2010 found a 87.7%-87.9% nucleotide homology and an over 98.8% amino acid homology. Conclusion The JEV isolated from the mosquitoes in Dinghai district and Yiwu city between year 1

关 键 词：脑炎病毒 日本 基因型 序列同源性 

分 类 号：R51[医药卫生—内科学] R3[医药卫生—临床医学]