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目的分析贵州省贵阳市2013—2015年手足口病患儿肠道病毒71型(EV71)VP1和VP4基因特征,为手足口病疫苗的选用和临床诊治提供参考依据。方法采集2013年8月—2015年4月在贵阳市第五人民医院就诊的120例手足口病轻症患儿鼻咽拭子和(或)粪便标本进行EV71病毒VP1和VP4全序列的逆转录-聚合酶链反应(RT-PCR)扩增和测序,并与美国国立生物技术信息中心公布的EV71 A、B、C基因型代表株进行系统进化分析,根据进化分析结果对此2个片段随机抽取20例与其他地区相同基因型进行氨基酸位点变异分析。结果所有样本病毒株均为C4基因亚型,与2008年北京、上海、安徽以及2011年深圳分离株成簇,20例EV71病毒株在VP1和VP4氨基酸位点变异与以上城市流行毒株相比,其中12例样本EV7的VP1片段氨基酸序列293位丙氨酸(A)变异为丝氨酸(S),其余位点均只有1例样本发生A26T、N31D、N282S突变,而VP4片段只有1个样本发生K52R突变。结论贵阳市2013—2015年手足口病患儿的EV71流行株均属C基因型的C4亚型,VP1片段常见A293S、A26T、N31D和N282S突变,VP4片段偶有K52R突变。
Objective To analyze the characteristics of VP1 and VP4 gene of enterovirus 71 (EV71) in children with hand-foot-mouth disease from 2013 to 2015 in Guiyang, Guizhou Province, and to provide reference for the selection and clinical diagnosis and treatment of HFMD. Methods A total of 120 VP1 and VP4 reverse transcripts of EV71 virus from 120 nasopharyngeal swabs and / or stool specimens of 120 children with HFMD were collected from August 2013 to April 2015 in the Fifth People’s Hospital of Guiyang. (PCR) amplification and sequencing, and phylogenetic analysis was performed with representative strains of EV71 A, B and C genotypes published by the National Center for Biotechnology Information. According to the results of evolutionary analysis, the two fragments were randomly selected A total of 20 cases of the same genotypes were selected for amino acid site variation analysis. Results All the sample strains were subtypes of C4 genotype, which were clustered with the isolates of Beijing, Shanghai, Anhui and Shenzhen of Shenzhen in 2008 and the variation of VP1 and VP4 amino acid positions in 20 EV71 strains compared with the above urban strains , Among them, the amino acid sequence of alanine at amino acid position 293 of 12 EV7 samples was changed to serine (S). Only 1 sample of other sites had A26T, N31D and N282S mutations, while only 1 sample of VP4 fragment K52R mutation. CONCLUSION: EV71 epidemic strains in children with HFMD in 2013-2015 in Guiyang belong to the C4 subtype of C genotype. A293S, A26T, N31D and N282S mutations are common in VP1 and K52R mutations in VP4.