2008年余姚市肠道病毒71型检测与VP1基因特征分析

VP1 gene analysis of enterovirus 71 strains isolated in 2008,Yuyao city

目的研究2008年余姚手足口病主要病原肠道病毒71型（EV71）毒株的分子生物学特征。方法收集手足口病患者粪便样品,用荧光逆转录-聚合酶链反应（RT-PCR）检测EV71,选择5个代表性样品进行VP1基因的扩增和核苷酸序列测定,测序结果利用DNAstar软件进行比对分析。结果 52例手足口病患者大便样品中检测到EV71阳性,阳性率为69.3%。5个代表株与A、B和C基因型标准株VP1基因核苷酸同源性分别为81.9%~82.2%、83.7%~84.3%和94.9%~95.3%,5个毒株之间的核苷酸同源性波动于96.2%~99.9%。在基因系统发生树上,这5个毒株均属于C基因型。结论本研究表明引起2008年余姚手足口病的主要病原是EV71毒株,它属于C基因型。

Objective To study VP1 gene of enterovirus 71（EV71） strains which were the major pathogen of hand-foot-mouth disease epidemics in Yuyao city in 2008.Methods The stool specimens were collected from the patients of hand-foot-mouth disease.EV71 infection was tested for all the specimens with a fluorescent reverse transcription polymerase chain reaction（RT-PCR）.The VP1 gene of EV71 was cloned from 5 representative samples and sequenced by DNAstar.Results EV71 was detected in 52 samples with a positive rate of 69.3%.The VP1 gene of Yuyao strains had 96.2%-99.9% sequence homologies with each other,and shared 81.9%-82.2%,83.7%-84.3% and 94.9%-95.3% sequence homologies with standard strain of A,B and C genotypes respectively.The Yuyao strains belonged to genotype C of EV71 in the phylogenetic tree.Conclusion The major pathogen of hand-foot-mouth disease in Yuyao city was EV71,which belonged to genotype C.