2010年广州地区儿童肠道病毒71型感染的分子流行病学研究

Molecular epidemiology study on human enterovirus 71 virus infection in Guangzhou in 2010

目的了解广州地区2010年肠道病毒(EV)71病毒流行状况。方法设计肠道病毒71型实时荧光RT-PCR引物和TaqMan探针,运用TaqMan实时荧光RT-PCR技术,对2010年全年来医院就诊的疑似手口足病患儿送检标本20150份,进行肠道病毒71型荧光RT-PCR检测,同时,设计了扩增EV71病毒全基因组的引物,随机选取了5株病毒进行全基因组进行测定。结果在20150份临床标本中,EV71病毒的阳性为4780份,全年总的检出率为23.72%,每个月均有阳性检出;其中,EV71病毒阳性率较高的为4～7月份,最高检出率为6月份,达32.19%,最低检出率为10月份,为7.91%;扩增了5株广州株EV71病毒全基因组全长序列,提交到GenBank上,将5株广州株EV71病毒与EV71的A、B3、B4、B5、C1、C2、C3、C4型及阜阳株全基因组核苷酸序列进行ClustalW比较,发现与C4a型阜阳株等同源性为98.00%～99.00%,与C4b同源性为91.00%～93.00%,与C1-C3同源性为82.00%～83.00%,与B3、B4、B5同源性为81.00%～83.00%,与A型同源性为80.00%。结论 2010年广州地区手足口病的主要病原是肠道病毒71型,EV71病毒在2010年全年还是较普遍流行,全基因组序列分析表明,2010年分离的5株广州株EV71病毒属于C4a型。

OBJECTIVE To study the molecular epidemiology of Human enterovirus 71 in Guangzhou in 2010 full year.METHODS Developed real-timeTaqMan RT-PCR for rapid detection of EV71 virus.A total 20150 clinical specimens were obtained from pediatric patients suspected HFMD in 2010 r.Primers were designed and amplified EV71 genome by RT-PCR.RESULTS Totally 4780 human entervirus 71 were indentified from 20150 samples and carrying rate of EV71 virus was 23.72%,relatively high positive rates of EV71 virus were Apr,May,Jun and Jul,the highest positive rate of EV71 virus in 2010 was Jun,with 32.19%,the lowest detection rate of EV71 virus in 2010 was Oct month,with 7.91%.The 5 strains EV71 genomic sequences are amplified and sequenced.The genomic sequences of EV71 Guangzhou strains were compared with those of EV71 in GenBank,revealed that the homology with EV71 genotype C4a Fuyang strains ranged between 98.00%-99.00%,homology with genotype C4b were 91.00%-93.00%,with genotypes C1,C2 and C3 were 82.00%-83.00%,with genotypes B3,B4,B5 were 81.00%-83.00%,homology with genotype A was 80.00%.CONCLUSION The main causative rates of EV71 virus is still higher in 2010 full year.Sequences and phylogenetics analysis of those Guangzhou strains in 2010 reveals that those isolates belongs to genotype C4a.agent of hand,foot and mouth disease（HFMD） in Guangzhou is EV71 virus in 2010.