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作 者:郑官增[1] 李桂霞[1] 裘丹红[1] 沈伟伟[1] 朱智勇[2]
机构地区:[1]浙江省台州市疾病预防控制中心,浙江台州318000 [2]浙江省疾病预防控制中心,杭州310051
出 处:《中国卫生检验杂志》2011年第11期2612-2615,共4页Chinese Journal of Health Laboratory Technology
摘 要: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.
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