2013年杭州地区肠道病毒71型分离株VP1基因特征分析  

Genetic analysis of the VP1 gene of enterovirus 71 isolated from Hangzhou in 2013

作　　者：宋闻 林先耀 王娟 吴亦栋 蒋茂莹 赵仕勇 

机构地区：[1]杭州市儿童医院感染科,310014

出　　处：《国际流行病学传染病学杂志》2015年第2期80-85,共6页International Journal of Epidemiology and Infectious Disease

基　　金：浙江省医药卫生科技计划项目（2012KYB159）;浙江省科技计划项目（2011F20075）

摘　　要：目的了解2013年杭州地区重症手足口病患儿感染的肠道病毒71型（EV71）分离株VP1区的基因特征。方法采集2013年杭州市儿童医院收治的重症手足口病确诊病例粪便样本，用EV71特异性引物进行RT·PCR鉴定，选取EV71检测阳性的15例样本进行病毒分离并进行VP1基因的特异性扩增并测序，结合EV71各基因型及亚型的代表株构建系统进化树，并对VP1蛋白的氨基酸序列进行分析。结果2013年该院分离的15株EV71分离株与A、B基因型参考株的核苷酸同源性分别为77．5％-79．8％、77．7％-83．1％，氨基酸同源性分别为9218％-96．6％、92．8％-97．9％，与C基因型的核苷酸同源性为83．4％-97．3％，氨基酸同源性为93．4％-99．7％，其中与2008年安徽省阜阳市的EV71流行株[C4a（Fuyang-0508）]的核苷酸和氨基酸同源性最高，分别为95．0％-97.3％及96．2％-99．7％。系统进化树显示所有分离株与C4a亚型的代表株处于同一分支。15株EV71分离株与不同基因型代表株进行比对发现，在297个氨基酸位点中主要突变位点有11个。结论2013年杭州地区重症手足口病患儿中流行的EV71均为C4a基因型，其VP1基因保守性好。Objective To investigate the genetic characteristics of the VP1 gene, enterovirus 71 （EV71） of isolated strains in Hangzhou in 2013. Methods To collect the stool specimens of the confirmed cases of severe HFMD from Hangzhou Children＇s Hospital in 2013.All of the specimens were identified by reverse transcription polymerase chain reaction （RT-PCR） with EV71-type-specific primers. Full length VP1 genes of EV71 isolated strains from 15 positive specimens were specifically amplified by RT-PCR and sequenced. The sequences were compared with VP1 genes from EV71 in GenBank database. The amino acid sequence of VP1 protein was analyzed. Results The nucleotide homogeneities between these 15 Hangzhou strains and the representative isolates of genotype A and B were 77.5%-79.8% and 77.7%-83.1%, respectively; while the amino acid homogeneities were 92.8%-96.6% and 92.8%-97.9%, respectively. The nucleotide and amino acid homogeneities between the 15 Hangzhou strains and the representative isolates of genotype C were 83.4%-97.3% and 93.4%- 99.7%, respectively. The 15 Hangzhou strains had the closest homology with Cga （Fuyang-0508） strain in 2008, of which the nucleotide and amino acid homogeneities appeared to be 95%-97.3% and 96.2%-99.7%, respectively. The 15 EY71 Hangzhou strains were at the same branch as the representative strain of subgenogroup Cga in tile phylogenetic tree. There were 11 main point mutations from the total 279 amino acid points between the 15 Hangzhou strains and other representative strains. Conclusions The EV71 strains isolated from severe HFMD of Hangzhou in 2013 belong to subgenogroup C4a, and the VP1 gene is conservative.

关 键 词：肠道病毒感染 VP1基因 序列分析 点突变 

分 类 号：R725.1[医药卫生—儿科]