宁波市肠道病毒71型分离株基因组特征研究

Research on the genomic characteristics of Enterovirus 71 isolates in Ningbo area

目的:了解2010年宁波市分离的肠道病毒71型的遗传进化来源。方法:设计八对引物用RT-PCR扩增、拼接得到EV71基因组序列,利用DNA-STAR对结构蛋白VP1-VP4和三个前体蛋白P1、P2及P3进行遗传进化分析。结果:宁波株NB10基因组核苷酸序列长度为7406 bp,与北京08年分离株FJ606448基因组间的同源性最高(高达99%),两者结构蛋白氨基酸序列完全相同。NB10与C4亚型的其他参考株,如:安徽株(GQ994988 FY08)、深圳株(FJ607337.1SHEZH08)、等核苷酸同源性均大于98%,而与标准株(BrCr)和(MS-7423-87)的核苷酸的进化关系较远,在P1、P2、P3区,NB10与C4亚型各代表株的同源性均为94%以上;而与北京09株(FJ606450.1)在P3区的核苷酸序列和推导的氨基酸序列之间同源性均较低,分别为87.9%和96.7%。结论:宁波株NB10与中国大陆北京、阜阳、深圳等毒株亲缘关系较近,而与北京株(FJ606450.1)有较大的差异,可能为不同的进化来源。

Objective：To understand the genetic evolution origin of Enterovirus 71 strains isolated in Ningbo area in 2010.Methods：The genome were amplified and sequenced with eight fragments with RT-PCR.Then the amino acid sequences and nucleotide sequences of VP1,VP2,VP3,VP4,P1,P2 and P3 sequences were analyzed by DNA-STAR.Results：The full-length of NB10 strain was 7406bp,had the most similarities with the strains FJ606448 isolated in Beijing in 2008.The similarity was 99%.The Amino acid sequences of the Structural protein were the same between the two strains.The similarities of genomes between strain NB10 with other EV71 stains of C4 subtype such as Anhui strain CQ994988 FY08,Shenzhen strain FJ607337.ISHEZH08,were all more than 98%,while the similarities between NB10 and the standard stains BrCr and MS-7423-87 were far.In area of p1,p2 and p3,the similarities between NB10 and other strains of C4 subtype were more than 94%.Only in P3 area,the similarities of amino acid sequences and nucleotide sequences between NB10 and Beijing strain FJ606450.1 were 96.7% and 87.9%.Conclusion：The genetic relationship between NB10 and other reference strains of C4 subtype from mainland was near,while the genetic relationship between NB10 and Beijing strain FJ606450.1 was far.Maybe NB10 and Beijing strain FJ606450.1 origined from different EV71 strains.