辽宁省乙脑病毒的分离与鉴定  

Isolation and Identification of Japanese encephalitis virus in Liaoning Province

作　　者：王俊文[1] 付士红[2] 王环宇[2] 毛晓燕[3] 刘卫滨[2] 何英[2] 孙肖红[2] 蔡增林[4] 吴立平[5] 赵习芳[5] 韩瑞红[6] 景雅[1] 梁国栋[2] 

机构地区：[1]山西医科大学组胚教研室,太原030001 [2]中国疾病预防控制中心病毒病预防控制所 [3]兰州生物制品研究所 [4]沈阳军区军事医学研究所 [5]山西疾病预防控制中心 [6]中国医学科学院实验动物研究所

出　　处：《中华实验和临床病毒学杂志》2006年第1期61-65,F0003,共6页Chinese Journal of Experimental and Clinical Virology

基　　金：科技部重大传染病专项(2003BA712A08-01)

摘　　要：目的 对2002年辽宁省采集的蚊虫标本进行病毒分离与鉴定。方法 从辽宁省采集蚊虫标本4927只，用细胞培养的方法分离病毒，对新分离的病毒进行血清学（ELISA和IFA方法）、分子生物学鉴定。结果 本实验共研磨蚊虫标本50批，分离到2株病毒，命名为LN02-102、LN02，104。这2株病毒均可致BHK-21细胞病变（CPE）（2—3d），致Vero细胞病变（2—4d），也可致C6／36细胞病变（2—4d）。对3日龄乳鼠2—3d可致死。这2株病毒可与标准乙脑病毒抗体反应。利用病毒PrM区段（基因组456-695核苷酸）进行基因分型分析。新分离的2株病毒属于基因Ⅰ型乙脑病毒。对这2株病毒的E基因区段进行分析，它们之间核苷酸和氨基酸的同源性为100％，与疫苗株SA14-14-2相比，核苷酸差异为4．11％，氨基酸差异在0．60％。结论 在辽宁省采集的蚊虫标本中分离到2株病毒，经血清学和分子生物学鉴定为基因Ⅰ型乙脑病毒，是近年来在辽宁省首次分离到基因Ⅰ型的乙脑病毒。Objective To study arboviruses carried by mosquitoes in Liaoning Province in 2002. Methods Totally 4927 mosquitoes were collected from Liaoning Province in July 2002. Virus strains were isolated by inoculating BHK-21, C6/36 and Vero cells. The newly isolated strains were identified by serological （ELISA and IFA）and molecular methods （Real-Time PCR and RT-PCR）. Results Two strains were isolated from mosquitoes causing cytopathogenic effect （CPE） on cells and were fatal, to suckling mice. Serological tests showed that both were positive for the antibody to JEV. The PrM and E gene were cloned and sequenced. The phylogenetic analysis showed that the new isolates belonged to genotype Ⅰ , JEV. Sequence analysis showed that the homology of nucleotide sequences and amino acid （AA） sequences between the two strains was 100%. Compared with the nucleotide sequences between the two strains and the standard JEV vaccine strain SA14-14-2, the difference was up to 4.11%, and the difference of AA was 0.6 %. Conclusion Two strains of JEV were isolated and identified in Liaoning province,both belonged to genotype Ⅰ JEV.

关 键 词：脑炎病毒 日本 种系发生 基因 

分 类 号：R373[医药卫生—病原生物学]