临床手足口病肠道病毒71型VP1基因分析  

Analysis of VP1 Gene of Enterovirus Type 71 from Clinical Hand,Foot,and Mouth Disease

作　　者：王杨[1] 李祎[1] 彭传梅 张胜[1] 陈婉婷[1] 卫波[1] 高辉[1] WANG Yang;LI Yi;PENG Chuarrmei(Department of Clinical Laboratory ,Yan ＂an Hospital Affiliated to Kunming Medical University, Kunming 650051 ,China)

机构地区：[1]昆明医科大学附属延安医院检验科,云南昆明650051

出　　处：《中国实验诊断学》2018年第5期811-814,共4页Chinese Journal of Laboratory Diagnosis

基　　金：云南省应用基础研究项目(2014FB080);云南省应用基础研究-昆医联合专项(2014FZ004)

摘　　要：目的对临床手足口病(HFMD)的病原体肠道病毒71型(EV71)分离株VP1基因测序分析,从分子水平探究HFMD的分子遗传进化变迁情况。方法收集2015年1月至2016年12月期间在延安医院临床就诊疑似HFMD患者590例,采集粪便标本,用实时荧光PCR法进行EV71病毒核酸检测,对EV71检测阳性粪便样本进行逆转录-聚合酶链反应(RT-PCR)扩增VP1目的基因,扩增阳性产物进行测序和序列比对,分析同源性并确定其所属基因亚型,并构建种系遗传进化发育树。结果共检出EV71型50例,检出率为8.47%(50/590例),2015年检出28例,2016年检出22例。50例中≤5岁35例,5-18岁13例,≥18岁2例;男性占64.00%(32/50例),女性36.00%(18/50例),女性患者中包括2例成人HFMD患者。检出的50株EV71毒株均为EV71C4亚型。检出的EV71毒株与国内北京和安徽以及马来西亚地区的毒株遗传距离为0.026,与浙江、广东地区毒株的遗传距离为1.52,与澳大利亚地区毒株的遗传距离为2.11。结论亲缘关系进化树显示,EV71毒株2年间未发生大的基因遗传变异。EV71毒株与北京、安徽以及马来西亚EV71流行毒株遗传距离较近,与浙江、广东等地的EV71流行毒株遗传距离较远,与澳大利亚分离的EV71毒株遗传距离最远。Objective The VP1 gene of pathogenic enterovirus 71（EV71）strain isolated from clinical hand-footmouth disease（HFMD）was sequenced to investigate the molecular genetic evolution of HFMD at the molecular level.Methods A total of 590 HFMD cases were collected from Yan＇an Hospital during Jan.2015 to Dec.2016.Fecal samples were collected.Real-time PCR was used to detect the EV71 virus nucleic acid.The VP1 genes of EV71 positive stool samples were amplified by RT-PCR.The amplified positive products were sequenced and aligned.The homologies were identified and their subgenotypes were determined.The phylogenetic tree was constructed.Results A total of 50 cases of EV71 were detected,with a detection rate of 8.47%（50/590 cases）,of which 28 cases were detected in 2015 and 22 cases were detected in 2016.Of the 50 cases,35 cases were≤5 and 5-18 years old cases;≥18 years old in 2 cases;male accounted for 64.00%（32/50 cases）,female 36.00%（18/50 cases）,and female patients including 2 cases of adult HFMD patients.All the 50 EV71 isolates were EV71 C4 subtypes.The genetic distance between detected strains of EV71 and the strains in Beijing,Anhui and Malaysia was 0.026.The genetic distance to the strains in Zhejiang and Guangdong was 1.52,and to the strains in Australia was 2.11.Conclusion Phylogenetic tree showed that there was no major genetic variation in the EV71 strains in two years.EV71 isolates were close to those of epidemic strains in Beijing,Anhui and Malaysia.They were far from the epidemic strains in Zhejiang and Guangdong,and the most distant from the strains in Australia.

关 键 词：手足口病 肠道病毒71型 VP1基因 DNA测序 系统发育树 

分 类 号：R725.1[医药卫生—儿科]