云南省5株柯萨奇病毒A组9型进化分析  

Phylogenetic analysis of 5 strains of coxsackievirus A9 isolated from Yunnan province

作　　者：赵丽娟[1] 姜黎黎[2] 周晓芳[2] 寸建萍[2] 伏晓庆[2] 田炳均[2] ZHAO Lijuan;JIANG Lili;ZHOU Xiaofang;CUN Jianping;FU Xiaoqing;TIAN Bingjun(Baoshan Center for Disease Control and Prevention,Baoshan,Yunnan 678000,China;不详)

机构地区：[1]保山市疾病预防控制中心急性传染病科,云南保山678000 [2]云南省疾病预防控制中心急性传染病防制所,云南昆明650022

出　　处：《中国病毒病杂志》2023年第5期378-383,共6页Chinese Journal of Viral Diseases

基　　金：国家“十三五”艾滋病和病毒性肝炎等重大传染病防治科技重大专项(2017ZX10103010)。

摘　　要：目的对云南省手足口病(hand,foot and mouth disease,HFMD)病例中检测到的柯萨奇病毒A组9型(coxsackievirus A9,CVA9)VP1区基因进行基因进化分析,为今后CVA9的预防及控制提供实验室依据。方法对2014―2021年云南省疾病预防控制中心手足口病实验室收到的手足口病儿童粪便标本悬液提取病毒RNA,先用MD91/OL68-1引物扩增肠道病毒(enterovirus,EV)VP4/VP2结合区基因并测序,编辑后的序列在GenBank上进行“BLAST”搜索确定病毒型别,再用肠道病毒B组引物扩增CVA9病毒VP1区基因全序列并测序,通过肠道病毒在线定型工具(Enterovirus Genotyping Tool Version 0.1)进行病毒型别确认。在基因库(GenBank)下载CVA9病毒VP1区基因型/亚型代表株序列,用Mega 5.2软件构建系统进化树,分析病毒基因特征。结果2014―2021年,云南省手足口病实验室共从手足口病病例粪便标本中检测到5株CVA9病毒。基因特征分析结果表明,云南省5株病毒为基因型D中D2亚型的分支1,跟大多数中国分离株一致,是目前为止中国最为流行的基因亚型。结论云南省检测到的5株CVA9病毒,是手足口病的稀有病原。加强CVA9病毒的实验室监测特别是分子流行病学监测,对追踪和分析病毒传染来源具有重要意义。Objective To conduct phylogenetic analysis of the VP1 gene of coxsackievirus A9(CVA9)isolated from hand,foot and mouth disease(HFMD)cases in Yunnan province,in order to provide laboratory evidence for prevention and control of CVA9 in the future.Methods Stool suspensions were made using the stool samples from children HFMD cases submitted to the HFMD Laboratory at Yunnan Center for Disease Control and Prevention in the period from 2014 to 2021.Enterovirus(EV)RNA was abstracted directly from the treated stool suspensions.EV VP4/VP2 junction region sequences were firstly amplified by MD91/OL68-1 primer pairs and sequenced.The edited sequences were queried using BLAST in the GenBank to determine the virus serotype.To analyze the phylogenetic characteristics of CVA9,the entire VP1 gene sequences were amplified using EV species B(EV-B)primer and sequenced.The serotype was confirmed using the online“Enterovirus Genotyping Tool Version 0.1”.The sequences of the reference genotypes/sub-genotypes of CVA9 VP1 region were downloaded from GenBank.The phylogenetic trees were constructed by Mega 5.2 software and the genetic characteristics were analyzed.Results From 2014 to 2021,a total of 5 strains of CVA9 were detected in the stool samples from HFMD patients in Yunnan provincial HFMD laboratory.Phylogenetic analysis showed that CVA9 could be divided into 4 genotypes,namely A,B,C and D.Similar to most isolates in China,the 5 strains isolated in Yunnan province all belonged to cluster 1 of sub-genotype D2,the dominant sub-genotype circulating in China so far.Conclutions The 5 strains of CVA9 detected from HFMD network in Yunnan province is a rare pathogen for HFMD patients.It is important to strength the laboratory surveillance,especially molecular epidemiological surveillance,for tracking and analyzing transmission source of CVA9.

关 键 词：柯萨奇病毒A组9型 手足口病 VP1区基因 基因特征分析 

分 类 号：R373.2[医药卫生—病原生物学]