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目的明确邯郸市2010-2012年EV71流行株的基因型及其VP1区基因特征,为手足口病防治提供分子生物学依据。方法用RD细胞分离培养EV71病毒,RT-PCR方法扩增VP1区基因片段,进行核苷酸和氨基酸序列同源性分析,构建系统进化树。结果邯郸市EV71分离株VP1区核苷酸同源性为96.4%~100.0%,与A、B基因型代表株VP1区核苷酸同源性在81.8%~82.7%与83.6%~84.5%之间。与已知基因型C1、C2、C3、C4代表株的核苷酸同源性分别在89.9%~90.3%、89.3%~90.1%、87.9%~88.7%和96.0%~96.9%之间,与1998年台湾分离株同源性为92.4%~93.2%,与2008年阜阳、2009年河南、2010年河北和2011年陕西分离的EV71代表株同源性在96.6%~99.1%之间。基于VP1区构建的EV71系统进化树显示,这14株EV71均为C4a亚型。结论邯郸市2010-2012年EV71流行株为C4a亚型,与中国大陆近年流行株分离结果一致。
Objective To identify the genotypes and VP1 gene characteristics of EV71 strains from 2010 to 2012 in Handan City and to provide a molecular basis for the prevention and treatment of HFMD. Methods The EV71 virus was isolated and cultured by RD cells. The VP1 gene fragment was amplified by RT-PCR. The nucleotide and amino acid sequence homology was analyzed to construct the phylogenetic tree. Results The nucleotide homology of VP1 of Handan EV71 isolate was 96.4% -100.0%, and the nucleotide homology of the VP1 gene of strains A and B was 81.8% -82.7% and 83.6% -84.5% between. Nucleotide homology with the representative strains of C1, C2, C3 and C4 was 89.9% -90.3%, 89.3% -90.1%, 87.9% -88.7% and 96.0% -96.9%, respectively, with The isolates from Taiwan isolated in 1998 were 92.4% -93.2% homologous with the strains of EV71 isolated from Fuyang in 2008, Henan in 2009, Hebei in 2010 and Shaanxi in 2011, and their homologies ranged from 96.6% to 99.1%. The EV71 phylogenetic tree based on the VP1 region showed that all 14 EV71 were C4a subtypes. Conclusion The EV71 epidemic strain in the city of Handan from 2010 to 2012 was C4a subtype, which was consistent with the isolation of the epidemic strains in mainland China in recent years.