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目的:研究2010年肠道病毒71型宁波分离株的全基因组序列特征。方法:收集宁波市手足口病患者的粪便标本进行特异性荧光定量RT-PCR鉴定,EV71阳性标本进行病毒分离和全基因组序列的测定。结果:2010NB65株全长7406 bp,与参考株EV71相比编码区没有核苷酸的缺失和插入。2010NB65株与2008年-2010年中国大陆EV71流行株的各区段核苷酸和氨基酸同源性均较高,分别为94.1%～98.8%和97.6%～100%;与A型和CoxA16核苷酸同源性较低,分别为75.0%～82.4%和63.0%～85.4%。VP1和全基因组亲缘进化分析表明,2010NB65株属于C4基因亚型的C4a进化分支,与浙江株、阜阳株、北京株亲缘关系近。2010NB65株P2、P3区与CoxA16的进化途径相近。结论:宁波市HFMD患者粪便中分离得到EV71属于C4基因亚型的C4a进化分支,与我国其他地区的EV71存在共同进化趋势,全基因组序列提供了完整的分子生物学背景,为EV71病毒性疾病的防控提供参考价值。
Objective: To study the whole genome sequence of enterovirus 71 Ningbo isolate in 2010. Methods: Stool specimens of hand, foot and mouth disease patients in Ningbo were collected for specific fluorescent quantitative RT-PCR and EV71 positive samples for virus isolation and genome-wide sequence determination. Results: 2010NB65 strain was 7406 bp in length. Compared with the reference strain EV71, there was no nucleotide deletion and insertion in the coding region. The nucleotide and amino acid homology of 2010NB65 strain and the EV71 epidemic strain in mainland of China from 2008 to 2010 were high, which ranged from 94.1% to 98.8% and 97.6% to 100%, respectively. Compared with A and CoxA16 nucleotide The homology was low, ranging from 75.0% to 82.4% and 63.0% to 85.4%, respectively. Phylogenetic analysis of VP1 and whole genome showed that 2010NB65 belonged to the C4a evolutionary branch of C4 subtype, which was closely related to Zhejiang strain, Fuyang strain and Beijing strain. 2010NB65 strains P2, P3 and CoxA16 evolutionary pathway similar. Conclusion: EV71 belongs to the C4a evolutionary branch of C4 subtype in feces of patients with HFMD in Ningbo City. It has a common evolutionary trend with EV71 in other regions of China. The complete genome sequence provides a complete molecular biology background for EV71 viral diseases Prevention and control provide a reference value.