柯萨奇病毒A组9型的基因分型方法的建立  

Establishment of A Genotyping Method for Coxsackievirus A9

作　　者：赵鹤鹤 冀天娇[1] 杨倩[1] 肖金波 祝双利[1] 王东艳[1] 王建醒[2] 陈建华[3] 路环环 韩振志 张勇[1,4] 许文波[1,4] 严冬梅[1] ZHAO Hehe;JI Tianjiao;YANG Qian;XIAO Jinbo;ZHU Shuangli;WANG Dongyan;WANG Jianxing;CHEN Jianhua;LU Huanhuan;HAN Zhenzhi;ZHANG Yong;XU Wenbo;YAN Dongmei(National Polio Laboratory,WHO WPRO Regional Polio Reference Laboratory,National Health Commission Key Laboratory for Biosecurity,National Health Commission Key Laboratory for Medical Virology,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 1022206,China;Shandong Center for Disease Control and Prevention,Jinan,250014,China;Gansu Center for Disease Control and Prevention,Lanzhou,730000,China;Center for Biosafety Mega-Science,Chinese Academy of Sciences,Beijing 102206,China)

机构地区：[1]国家脊髓灰质炎实验室,世界卫生组织西太平洋区脊髓灰质炎参比实验室,国家卫生健康委员会,生物安全重点实验室,国家卫生健康委员会,医学病毒和病毒病重点实验室,中国疾病预防控制中心,病毒病预防控制所,北京102206 [2]山东省疾病预防控制中心,济南250014 [3]甘肃省疾病预防控制中心,兰州730000 [4]中国科学院生物安全大科学研究中心,北京102206

出　　处：《病毒学报》2022年第2期264-270,共7页Chinese Journal of Virology

基　　金：北京市自然科学基金(项目号:L192014),题目:EVA71疫苗大规模应用后对我国手足口病病原谱的影响及其保护效果评价研究;国家重点研发计划(项目号:2021YFC0863000),题目:新型冠状病毒冷链传播机制和低温消毒技术研究。

摘　　要：血清型别鉴定及基因分型分析是开展肠道病毒(Enterovirus,EV)分子进化特征研究的重要内容。目前为止,国内外对柯萨奇病毒A组9型(Coxsackievirus A9,CVA9)的研究主要集中在衣壳蛋白区的细胞受体结合位点及其基因特性分析,而基于全长VP1序列的基因型划分结果尚未明确。本研究依托国家手足口病监测网络,对2010-2019年全国31个省级行政区(省、自治区、直辖市)上送的18 238份手足口病样本中分离出的24株CVA9进行全长VP1区序列测定,并与GenBank中所有全长VP1区序列一起进行基因型划分研究。测序结果显示24株CVA9分离株VP1全长为906bp,编码302个氨基酸,与CVA9原型株(Griggs)核苷酸和氨基酸相似性分别为80.5%~97.6%和92.3%~99.6%。结合系统进化树和同一血清型内不同基因型的核苷酸差异界值为15%~25%,将全球CVA9划分为A-H八个基因型。进化树显示B、C和D基因型在病毒进化过程中已消失,而E、F和G基因型呈现共循环的趋势,其中G基因型包含了亚洲、北美洲、大洋洲和欧洲等9个国家的毒株,是CVA9的优势基因型。大部分中国株属于G基因型,与世界流行趋势一致。但在2018年新疆一例手足口病患儿体内分离到的CVA9毒株却与俄罗斯有很近的亲缘关系,被划分为F基因型,推测该毒株为俄罗斯输入株。CVA9虽具有明显的地理聚性,但G基因型中包含的多国家和多时间段的毒株也提示CVA9病毒可能存在远距离的传播和流行。Serotype identification and genotype classification are important components of the study of the molecular evolution of enteroviruses. Studies on coxsackievirus A9(CVA9)have focused mainly on analyses of the receptor binding sites in the viral protein(VP)1 region and its genetic characteristics.Results of genotype classification based on full-length VP1 are not clear. This study was based on pathogen surveillance in hand,foot and mouth disease(HFMD)in China. Twenty-four strains of CVA9 from 31 provinces were sequenced and analyzed together with all the full-length VP1 sequences in GenBank. Nucleotide sequencing revealed that the full-length VP1 of 24 CVA9 was 906 bases encoding 302 amino acids. The similarity in nucleotides and amino acids with the prototype strain of CVA9(Griggs)was 80.5%-97.6% and 92.3%-99.6%,respectively.Combination of the thresholds of the phylogenetic tree and nucleotide difference of different genotypes within the same serotype led to a value of 15%-25%,and enabled CVA9 to be categorized into eight genotypes:A-H.The phylogenetic tree showed that the B,C,and D genotypes had disappeared during virus evolution,whereas the E,F,and G genotypes showed co-circulation. Lineage G was the dominant genotype of CVA9 and included most of the strains from nine countries in Asia,North America,Oceania and Europe. Most Chinese strains belonged to the G genotype,suggesting that the molecular epidemiology was consistent with that observed worldwide. However,one CVA9 strain isolated from a child with HFMD in Xinjiang in China in 2018 was closely related to Russia strains,and was categorized as the F genotype,and suggested that the strain was imported from Russia. Although CVA9 has obvious geographic clustering,the multi-country and multi-period strains in the G genotype suggest that there may be long-distance transmission and prevalence of CVA9 worldwide.

关 键 词：柯萨奇病毒A组9型(CVA9) 基因分型 分子进化 

分 类 号：R373.2[医药卫生—病原生物学]