手足口病EV71病毒株的分离鉴定及其生物学特性研究

Research on isolation,identification and biologic characteristics of EV71 strains in HFMD

目的:为了获得肠道病毒71型病毒株,同时验证Vero细胞分离肠道病毒的效果。方法:采集手足口病例疱疹液、咽拭子标本,接种单层Vero细胞,观察细胞病变(CPE);CPE阳性的培养物用荧光定量RT-PCR方法进行鉴定及分型,选取毒株进一步做EV71 VP1全序列测定、电子显微镜形态学观察、间接免疫荧光染色、病毒CCID50测定。结果:从23例手足口病例33份标本中,分离到22株肠道病毒,分离阳性率为66.7%;其中肠道病毒71型17株占阳性数的77.3%,CoxA16型4株占18.2%,未分型1株占4.5%,经基因序列分析表明本次分离的肠道病毒71型毒株均为C基因型。结论:Vero细胞分离肠道病毒阳性率高,获得了17株EV71型毒株。

Objective：To obtain EV71 virus strains and study the effect of isolating enterovirus with Vero cells.Methods： To observe the cytopathic effect（CPE） after the HFMD cases′ nasal swab and herpes were inoculated on Vero cells.The CPE positive culture were identified types by real-time RT-PCR.Some strains were selected to take further analysis： VP1 gene sequence,morphological observation of electron microscope,indirect immunofluorescence assay and the cell culture infective dose 50%（CCID50） determination.Results： 22 enterovirus strains were isolated from 33 samples,the positive rate was 66.7%,including 17 EV71（77.3%） strains,4 CoxA16（18.2%） strains and 1 unclassified（4.5%） strain.Conclusion： The effect of isolating enterovirus with Vero cells was good.17 EV71 strains can be acquired.