深圳口岸输入寨卡病毒的基因和生物学特性分析  

Genetic and biological characterization of Zika virus from human cases imported through Shenzhen Port

作　　者：王强[1] 杨扬[1] 郑海霞[1] 毕玉海[1,2,3] 宋敬东[4] 李力强 顾大勇 王培毅[1,7] 李世华[2] 刘升[2] 赵迎泽[2] 刘磊[1] 高福[1,2,3,4] 刘映霞[1] 

机构地区：[1]深圳市第三人民医院,深圳市病原微生物与免疫学重点实验室,深圳518112 [2]中国科学院微生物研究所病原微生物与免疫学重点实验室,北京100101 [3]中国科学院流感研究与预警中心,北京100101 [4]中国疾病预防控制中心病毒病预防控制所,北京102206 [5]深圳市华大基因研究院,深圳518083 [6]深圳检验检疫科学研究院,深圳518010 [7]Faculty of Biological Sciences,University of Leeds

出　　处：《科学通报》2016年第22期2463-2474,共12页Chinese Science Bulletin

基　　金：深圳市科技计划(ZDSYS201504301534057);深圳市"三名工程"资助

摘　　要：2016年2月14日下午,一名从斐济和萨摩亚旅游回国的旅客在深圳皇岗口岸入关时有发热症状,其在旅行期间有蚊虫叮咬史.经过病因筛查和诊断确诊为寨卡病毒感染.本研究利用乳鼠和细胞培养传代,成功从病人血清中分离到致病病原——寨卡病毒,命名为SZ_SMGC-1.分离病毒在电子显微镜下可见圆球形经典黄病毒特征,病毒颗粒直径大小约为50 nm.利用特异性引物扩增并获得了病毒全基因组序列,同源性分析显示：SZ_SMGC-1与我国输入性病例中斐济/萨摩亚来源病毒高度同源,同源性均为99.9%,与委内瑞拉来源病毒同源性在99.0%~99.4%之间.研究显示,寨卡病毒进化为亚洲和非洲2个分支.进一步遗传进化分析表明,SZ_SMGC-1毒株与近期我国分离毒株同属于亚洲分支;SZ_SMGC-1与斐济/萨摩亚来源病毒聚类在一个亚分支;委内瑞拉来源的病毒除我国首个病例处于单独的小分支外,其余病毒全部聚类到另一个小分支.生物学特性研究显示,SZ_SMGC-1寨卡病毒可以在蚊虫来源C6/36细胞和哺乳动物来源Vero,BHK-21细胞中生长,感染Vero和BHK-21细胞后可见明显病变并形成典型噬斑.值得注意的是,相同细胞系的不同细胞克隆株对寨卡病毒的敏感性不同.寨卡病毒感染敏感哺乳动物细胞系的建立,对病毒致病机制的深入研究及相关疫苗和药物的研发奠定了基础.寨卡病毒在我国的输入,给我国寨卡疫情防控带来了严峻挑战,必须提早采取灭蚊等防控措施,加紧对疫苗和相关药物的研发,做好预防寨卡流行的技术储备.During February to March 2016, a total of 13 Zika virus （ZIKV） cases were imported into China by travelers. Infection with ZIKV in humans results in fever, rash and has been associated with microcephaly as well as Guillain-Barr6 syndrome. ZIKV is a mosquito-borne flavivirus and Aedes mosquitoes are the main transmission vector species. Since Aedes are prevalent throughout China, ZIKV has the potential to become widely disseminated if not properly controlled. The genetic and biological characteristics of ZIKV should be thoroughly studied in order to enact effective countermeasures. A novel ZIKV strain （SZ_SMGC-1） was successfully isolated from the sera of an imported case returning to China from Fiji and Samoa at the Shenzhen Port. The entire viral genome was sequenced and compared to whole ZIKV genomes downloaded from GenBank. Results showed that SZ SMGC-1 possesses the highest nucleotide identity （99.9%） to ZIKV from Fiji and Samoa, and also has high nucleotide identity （99.0%-99.4%） to ZIKV from Venezuela. Additionally, ZIKV is know to have evolved into distinct Asian and African lineages, and our phylogenetic analyses showed that the ZIKVs isolated from all imported cases into China clustered into the Asian lineage, To study the biological characteristics of ZIKV, SZ_SMGC-1 was used to infect different cell lines including the mosquito- derived C6/36, monkey-derived Veto and hamster-derived BHK-21 cells. Infection with SZ_SMGC-1 in vitro resulted in considerable cytopathic effects （CPE） to mammalian cells and supported replication of the virus to high titers, but did not cause readily observable CPE in C6/36 cells despite virus replication to comparable titers with mammalian cell lines. Interestingly, two clones of Vero and BHK-21 cell lines were used in the present study, respectively, but ZIKV SZ_SMGC-1 replicated much quicker to high virus titers in one clone than the other. Furthermore, there are no effective vaccine or drug for ZIKV until now. The results of the genetic and biological

关 键 词：寨卡病毒 SZ_SMGC-1 基因组 遗传进化 生物学特性 细胞病变效应 

分 类 号：R373[医药卫生—病原生物学]