我国首次分离到基因Ⅰ型乙型脑炎病毒  

Isolation and identification of genotypeⅠJapanese encephalitis virus in China

作　　者：王环宇[1] 付士红[1] 李晓宇[1] 宋宏[1] 闵继光[2] 邓娟[1] 杨益良[1] Ichiro Kurane 梁国栋[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所 [2]上海中医药大学分部 [3]National Institute of Infection Diseases,tokyojapan1628640

出　　处：《中华微生物学和免疫学杂志》2004年第11期843-849,共7页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目 ( 3 0 170 0 46) ;日本国立传染性疾病研究所课题资助 (JapanHealthScienceFoundation)

摘　　要：目的　对新分离乙型脑炎病毒鉴定 ,了解病毒E基因区段的氨基酸序列特征及基因分型。方法　 2 0 0 1年在上海市采集蚊虫标本 ,用组织培养的方法分离到 7株病毒。进行病毒一般生物学鉴定 ,对病毒PrM和E基因区段约 2 0 0 0nt进行了扩增、克隆、测序。应用ClustalX软件做碱基配对和比较分析 ,种系发生采用PHYLIP软件包分析。结果　 7株病毒可以引起BHK 2 1细胞规律病变 ,病变时间为 6 0h。乳鼠脑内接种病毒后 72h死亡。所有新分离病毒与标准乙型脑炎病毒抗体阳性反应。用病毒PrM区段 (4 5 6～ 6 95位核苷酸 )进行的基因分型分析 ,新分离的 7株病毒属于基因Ⅰ型乙型脑炎病毒。以减毒活疫苗株SA14 14 2为标准 ,对 7株病毒的E基因区段进行分析 ,7株病毒之间核苷酸同源性在 97.7%以上 ,氨基酸同源性在 99.2 %以上 ;7株病毒与疫苗株SA14 14 2核苷酸差异在 12 .2 %左右 ,氨基酸差异在 2 .8%～ 3.2 %之间 ,共有 14个共同的氨基酸位点差异。结论　 2 0 0 1年在上海采集的蚊虫中新分离的 7株病毒属于基因Ⅰ型乙型脑炎病毒 。Objective To analysis the genotype and the character of amino acid(AA) sequences of the E protein of the newly isolated Japanese encephalitis viruses(JEV) from mosquitoes collected in Shanghai 2001. Methods Virus isolation was performed inoculating mosquito homogenates onto BHK-21. The isolated strains were identified by biological and molecular methods. The PrM and E gene of the newly isolated JEV strains were cloned and sequenced. Phylogenetic analysis was achieved by Phylip program. Two hundred and forty nucleotides sequences of PrM gene(map position 456-695) region in the new isolated JEV strains were selected and compared with the reference strains from GenBank, isolated from different countries, to do phylogenic analysis. Nucleotide sequence and AA sequence of the E gene were studied by bioinformatics software. Results Seven virus strains were isolated, with all causing cytopathogenic effect(CPE) in BHK-21 cells and fatal to suckling mice. Serological tests showed that all of these strains were positive to the antibody of JEV. The phylogenetic results showed that the newly isolates allbelonged to genotype Ⅰ, JEV. Sequence analysis showed that the homology of nucleotide sequence in these 7 strains was over 97.7% and that of AA sequence 99.2%. Compared with differences of the nucleotide sequence between these seven strains and the standard JE vaccine strain SA-14-14-2, were up to 12.2%, and differences of AA sequence ranged from 2.8% to 3.2%. Compared with the vaccine strain, there were 14 common AA variations in all the 7 strains. Conclusion Newly isolated JEV strains belonged to genotype Ⅰ, which was reported for the first time in China. [

关 键 词：乙型脑炎病毒 E基因 首次 SA14-14-2 分离 蚊虫 基因分型 种系发生 氨基酸序列 碱基配对 

分 类 号：R373[医药卫生—病原生物学] R512[医药卫生—基础医学]