2011—2020年福建省手足口病相关柯萨奇病毒A5型基因特征及重组分析  

Genetic characteristics and recombination analysis of hand,foot and mouth disease related coxsackievirus A5 in Fujian province from 2011 to 2020

作　　者：何文祥[1] 朱颖[1] 陈炜[1] 翁育伟[1] He Wenxiang;Zhu Ying;Chen Wei;Weng Yuwei(Fujian Provincial Key Laboratory of Zoonosis Research,Fujian Provincial Center for Disease Control and Prevention,Fuzhou 350001,China)

机构地区：[1]福建省疾病预防控制中心,福建省人兽共患病研究重点实验室,福州350001

出　　处：《中华实验和临床病毒学杂志》2022年第6期678-684,共7页Chinese Journal of Experimental and Clinical Virology

基　　金：国家科技重大专项课题(2017ZX10104001);福建省科技创新平台建设项目(2019Y2001);福建省卫生健康科技计划项目(2021GGB012);福建省卫生健康科技计划项目资助(2019-1-18)。

摘　　要：目的了解福建省手足口病(hand,foot and mouth disease,HFMD)相关柯萨奇病毒A5型(coxsackievirus A5,CV-A5)的基因特征及重组情况。方法对2011—2020年福建省各地市上送的HFMD阳性标本利用逆转录巢式聚合酶链反应(reverse transcription nested polymerase chain reaction,RT-nPCR)对CV-A5 VP1区进行扩增、测序并分型。采用人横纹肌肉瘤(rhabdomyosarcoma,RD)细胞分离CV-A5,对分离到的毒株进行二代测序以获取病毒全基因组序列。应用MEGA7.0进行系统进化分析。利用软件RDP4及SimPlot 3.5.1进行病毒基因重组分析。结果福建省2011—2020年HFMD相关CV-A5病例31例,以1~2岁幼儿(16/31)为主,男女比例为2.1∶1(21/10),含3例重症病例、1例死亡病例。27例病例的CV-A5分型鉴定成功,均属于E1基因亚型。获得6株CV-A5全基因组序列,与原型株Swartz的同源性为81.1%~81.6%。基于全基因组、P1、P2、P3区核苷酸序列的进化树分析结果显示,毒株2017FJFZ239为疑似重组株,在P2及P3区与2株湖北CV-A5参考株(GenBank登录号:MW079817、MN663160)分布在同一分支。RDP4结果显示,毒株2017FJFZ239与2株湖北CV-A5参考株(GenBank登录号:MW079817、MN663160)发生了相同的重组事件,可能的亲本序列来源于CV-A5参考株(GenBank登录号:KP289362)和CV-A2参考株(GenBank登录号:KX595284)。SimPlot重组分析显示,毒株2017FJFZ239在非结构蛋白2B、2C及P3区可能与CV-A2参考株(GenBank登录号:KX595284)发生重组。结论2011—2020年福建省HFMD相关CV-A5为E1基因亚型,存在基因重组毒株。Objective To investigate the gene characteristics and recombination of coxsackievirus A5(CV-A5)in Fujian province.Methods The VP1 regions of CV-A5 strains from positive samples of hand,foot and mouth disease(HFMD)collected from all cities in Fujian province from 2011 to 2020 were amplified by reverse transcription nested polymerase chain reaction(RT-nPCR),then sequenced and identified the subgenotypes.CV-A5 strains were isolated in rhabdomyosarcoma(RD)cells.The isolated virus strains were sequenced by next generation sequencing to obtain the whole genome sequences.MEGA7.0 was used for phylogenetic analysis.RDP4 and SimPlot 3.5.1 softwares were used to analyze viral gene recombination.Results Total thirty-one HFMD related CV-A5 cases were detected in Fujian province from 2011 to 2020,mainly in children aged 1 to 2 years(16/31),with a male/female ratio of 2.1∶1(21/10),including 3 severe cases and 1 deceased case.CV-A5 subgenotyping was successful in 27 cases,all of them belonged to E1 subgenotype.The whole genome sequences of six CV-A5 strains were obtained,and the homology with the prototype strain Swartz was 81.1%-81.6%.Phylogenetic analysis results based on the sequences of whole genome,P1,P2 and P3 regions showed that the virus strain 2017FJFZ239 is a suspected recombinant strain,which was distributed in the same branch as CV-A5 reference strains(GenBank accession number:MW079817 and MN663160)in P2 and P3 regions.Analyzing with RDP4 showed that the virus strain 2017FJFZ239 had the same recombination event with CV-A5 reference strains(GenBank accession number:MW079817 and MN663160)and the potential parental sequences were CV-A2(GenBank accession number:KX595284)and CV-A5(GenBank accession number:KP289362).Further analyzing with SimPlot showed that the virus strain 2017FJFZ239 may recombine with CV-A2 strain(GenBank accession number:KX595284)in non-structural protein 2B,2C and P3 regions.Conclusions From 2011 to 2020,HFMD related CV-A5 in Fujian province belongs to E1 subgenotype,and there is a recombinant stra

关 键 词：柯萨奇病毒A5型 系统进化分析 重组 

分 类 号：R373.23[医药卫生—病原生物学]