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目的:分析肠道病毒71型2010年宁波分离株的分子流行病学特征。方法:收集2010年宁波市手足口病患者临床标本177份,进行EV71荧光定量RT-PCR鉴定和病毒分离,采用RT-PCR对38株分离到的EV71进行VP1编码区基因扩增,并对扩增产物进行核苷酸序列测序和分析。根据VP1测序结果与国内外报道的各基因型和基因亚型EV71VP1序列进行同源性和亲缘进化分析。结果:88份鉴定为EV71的临床标本中共分离到38株病毒,分离株的VP1区核苷酸和氨基酸同源性分别是97.1%～100%和99.3%～100%。与C4a亚型的核苷酸和氨基酸同源性分别为93.2%～99.3%和98.3%～100%。亲缘进化树显示,宁波分离株全部属于C4基因亚型的C4a进化分支,并存在多个传播链。结论:宁波地区分离的EV71与近几年国内其他地区EV71分离株亲缘关系很近,有共同进化的趋势,属于C4基因亚型的C4a进化分支,并存在多个传播链。
Objective: To analyze the molecular epidemiological characteristics of enterovirus 71 Ningbo isolates in 2010. Methods: A total of 177 clinical samples of hand, foot and mouth disease were collected from Ningbo city in 2010 for EV71 quantitative RT-PCR and virus isolation. RT-PCR was used to amplify the VP1 coding region of 38 isolated EV71 strains. The product was sequenced and analyzed for nucleotide sequence. According to VP1 sequencing results, the homology and phylogenetic analysis of each genotype and subtypes EV71VP1 sequence reported in China and abroad were carried out. Results: A total of 38 viruses were isolated from 88 clinical samples identified as EV71. The nucleotide and amino acid homology of VP1 in isolates were 97.1% -100% and 99.3% -100%, respectively. The nucleotide and amino acid homologies with C4a subtypes were 93.2% -99.3% and 98.3% -100%, respectively. Phylogenetic tree showed that all isolates in Ningbo belonged to the C4a evolutionary branch of the C4 gene subtype and had multiple transmission chains. Conclusion: EV71 isolated from Ningbo is closely related to the EV71 isolates in other regions of China in recent years. It has a co-evolutionary trend and belongs to the C4a evolutionary branch of the C4 gene subtype with multiple transmission chains.