2016年山东省烟台地区手足口病主要病原谱及柯萨奇病毒A6型VP1区基因特征分析  

Research on pathogenic spectrum of hand,foot and mouth disease and genetic characteristics of VP1 of Coxsackievirus A6 in Yantai region,Shandong province,China in 2016

作　　者：孙振璐 牟晓东[2] 刘虹[3] 张瑞卿 徐迎春 宫连凤[2] 刘娟 高巧[2] 董兆静[2] 王佶 马学军[1,5] Sun Zhenlu;Mou Xiaodong;Liu Hong;Zhang Ruiqing;Xu Yingchun;Gong Lianfeng;Liu Juan;Gao Qiao;Dong ZhaoJing;Wang Ji;Ma Xuejun(NHC Key Laboratory of Medical Virology and Viral Diseases,National Institute for Viral Disease Control and Prevention,,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Yantai Center for Disease Control and Prevention,Yantai 264003;Yantai Qishan Hospital;Hebei Medical University,Shijiazhuang,050031,Hebei,China;Center for Biosafety Mega-science,Chinese academy of Science,Wuhan 430071,China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制研究所,国家卫生健康委员会医学病毒和病毒病重点实验室,北京102206 [2]烟台市疾病预防控制中心,264003 [3]烟台市奇山医院,264001 [4]河北医科大学,石家庄050017 [5]中国科学院生物安全大科学中心,武汉430071

出　　处：《中华实验和临床病毒学杂志》2020年第4期429-434,共6页Chinese Journal of Experimental and Clinical Virology

基　　金："十三五"国家科技重大专项课题(2018ZX10713002);2017-2018年度山东预防医学会智飞疾病预防控制技术研究基金项目计划(LYH2017-26)。

摘　　要：目的研究2016年山东省烟台地区手足口病(hand,foot and mouth disease,HFMD)的病原谱组成特点,分析烟台HFMD病原谱中柯萨奇病毒A组6型(coxsackie virus A group 6 type,CV-A6)流行株的进化及VP1区重要氨基酸位点的变异情况。方法采用real-time RT-PCR方法,对2016年烟台地区采集于HFMD患者的738份样品进行肠道病毒(enterovirus,EV)核酸检测和分子定型,统计各型EV阳性标本数量及比例。根据EV的优势型别,选取8株CV-A6进行VP1区扩增,并进行核苷酸序列测定和分析,然后进行系统进化分析。结果从738份标本中检出EV 460份,主要病原构成为柯萨奇病毒A组16型(coxsackie virus A group 16 type,CV-A16)(56.09%,258/460)、肠道病毒A组71型(enterovirus A group 71 type,EV-A71)(13.48%,62/460)、柯萨奇病毒A组10型(coxsackie virus A group 10 type,CV-A10)(10.65%,49/460)、CV-A6(9.57%,44/460)、柯萨奇病毒A组4型(coxsackie virus A group 4 type,CV-A4)(1.96%,9/460)。其中成功分离到8份CV-A6阳性标本毒株,并进行了全长VP1区核苷酸序列分析,8株CV-A6的序列分析结果表明,其核苷酸相似性为96.12%~100%,氨基酸相似性为97.78%~100%。系统进化分析显示8株CV-A6烟台株均属于D基因型中的D3基因亚型。与参考株CVA6-Gdula-AY421764相比,烟台市CVA16分离株VP1第10、14、174、194、279、283、305位氨基酸出现突变。结论CV-A16、EV-A71、CV-A10、CV-A6是引起2016年烟台地区HFMD发病的常见病原体,本次分离到的CV-A6均属于D基因型中的D3基因亚型。Objective To investigate the pathogenic spectrum of enteroviruses associated with hand,foot and mouth disease(HFMD)in the Yantai region of Shandong province in 2016,and analyze the evolution of epidemic strains of coxsackie virus group A type 6(CV-A6)in the pathogenic spectrum of HFMD enteroviruses and the variations of important amino acid sites in the VP1 region.Methods A total of 738 samples were collected from the patients with HFMD in Yantai region in 2016 to conduct DNA and serotype tests of enterovirus(EV)by real-time RT-PCR and further count the number and proportion of each type of enterovirus positive specimens.Based on the predominant serotype of enteroviruses,eight serotypes of the CV-A6 strains were selected to carry out VP1 regions amplification for the determination and analysis of nucleotide sequencing and phylogenetic analysis.Results A total of 460 enteroviruses strains were isolated from 738 samples,including pathogens strains:258 CV-A16(56.09%),62 EV-A71(13.48%),49 CV-A10(10.65%),44 CV-A6(9.57%)and 9 CV-A4(1.96%).Eight CV-A6 positive specimens were isolated from the viruses and the nucleotide-sequence analysis of the whole VP1 region was conducted.The sequence analysis of eight CV-A6 strains demonstrated that the homologies of nucleotide and amino acid were 96.12%-100%and 97.78%-100%respectively.The phylogenetic analysis indicated that the eight CV-A6 strains were subdivided into the genotype D subtype D3.Compared with the reference strain,CVA6-Gdula-AY421764,amino acids of CV-A6 strains in Yantai city observed at sites 10,14,174,194,279,283 and 305 in VP1 region appeared mutant.Conclusions CV-A16,EV-A71,CV-A10 and CV-A6 were the main common pathogens of HFMD in Yantai region in 2016.All the CV-A6 strains isolated in this study belonged to subtype D3 in genotype D.

关 键 词：手足口病 病原谱 柯萨奇病毒 系统进化分析 

分 类 号：R725.1[医药卫生—儿科] R440[医药卫生—临床医学]