江苏省2009—2013年汉坦病毒分离株全基因序列测定及分析

目的:从分子水平分析2009—2013年江苏省分离汉坦病毒(Hantavirus,HV)的型别及变化,为疾病的预防控制提供实验依据?方法:采集2009—2013年江苏省肾综合征出血热(hemorrhagic fever with renal syndrome,HFRS)急性期患者血清及监测点鼠肺标本?采用胶体金法检测血清样本中汉坦病毒的IgM?IgG抗体,采用免疫荧光检测鼠肺样本中汉坦病毒抗原,并用VERO E6细胞分离病毒,RT-PCR检测标本培养物中病毒M片段以鉴定HV病毒,采用高通量测序技术对分离株进行全基因组测序,MEGA6.0软件对其进行序列比对和进化分析?结果:分离到的11株病毒与陕西分离株LR1?四川分离株S85-46以及汉坦病毒原型株76-118遗传距离最为接近,属于汉滩(Hantean,HTN)型病毒?分离株与2002年分离的江苏(A9?HTN型)株以及浙江株?安徽株处于不同的进化分支上,表明遗传关系较远?结论:江苏的汉坦病毒优势株发生了变化,而且发生了宿主转换?目前疫苗仍具有保护效力,但需加强监测?

Gene sequence determination and analysis of Hantavirus isolated strains in Jiangsu province from 2009 to 2013

Objective:The gene sequence of Hantavirus isolated strains during 2009 and 2013 was analyzed at molecular level to provide evidence for the prevention and control of this disease. Methods:The serum of patients with acute hemorrhagic fever with renal syndrome(HFRS)and rat lung specimens were collected from 2009 to 2013. Hantavirus IgM and IgG antibody in serum samples and antigen in mouse lung samples were detected using colloidal gold and IFA assay,respectively. With VERO E6 cell line,Hantavirus was isolated,then analyzed using RT-PCR assays,and genome was sequenced using high-throughput sequencing technology. Finally,sequence alignment and phylogenetic analysis were accomplished with MEGA6.0 software. Results:With the closest genetic distance,isolated 11 virus isolates and Shanxi LR1,Sichuan isolates s85-46 and Hantavirus strain 76-118 prototype were all classified as Hantaan (HTN)virus. In addition,strains isolated in Jiangsu 2002 (A9,HTN type)and strains in Zhejiang and Anhui on different evolutionary branches,indicated a larger genetic distance. Conclusion:Both Hantavirus predominant strains switchover and host transfer have happened in Jiangsu. Surveillance should be strengthened to guarantee the efficacy of the vaccine.