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目的分析2009年北京市西城区手足口病柯萨奇病毒A5型(CoxA5)的VP4区基因特征,了解其变异情况。方法采集手足口病患儿咽试子,提取病毒RNA,用肠道病毒VP4区分型引物进行半巢式RT-PCR(RT-nPCR),通过测序和GenBank Blast比对确定肠道病毒型别,并对其VP4区进行序列和进化分析。结果共鉴定出5株CoxA5,GenBank登录号为JN981017-JN981021。在VP4区,5株CoxA5与中国株HQ728261核苷酸及氨基酸序列同源性分别为96%～99%和98%～100%。进化分析表明,5株CoxA5与中国株HQ728261遗传距离为0.005～0.045,共同构成一个独立的进化树分支。5株CoxA5在VP4区无氨基酸的缺失和插入,未发现特有的氨基酸置换。结论 2009年北京市西城区手足口病病原体CoxA5VP4区与中国株HQ728261在进化上密切相关,其序列未发生明显变异。
Objective To analyze the gene characteristics of VP4 region of CoxA5 in hand, foot and mouth disease in Xicheng District of Beijing in 2009 and investigate its variation. Methods The pharyngeal testis was collected from children with hand-foot-mouth disease and the viral RNA was extracted. The semi-nested RT-PCR (RT-nPCR) was performed with the VP4 differentiating type of enterovirus and the enterovirus type was determined by sequencing and GenBank Blast comparison. And its VP4 region sequence and evolutionary analysis. Results A total of 5 CoxA5 strains were identified and their GenBank accession numbers were JN981017-JN981021. In the VP4 region, the nucleotide and amino acid sequence homologies of the five CoxA5 and Chinese HQ728261 were 96% -99% and 98% -100%, respectively. Phylogenetic analysis showed that the genetic distances between the five CoxA5 strains and the Chinese strain HQ728261 were between 0.005 and 0.045, forming an independent branch of the phylogenetic tree. There were no amino acid deletions and insertions in the 5 strains of CoxA5 in the VP4 region, and no unique amino acid substitutions were found. Conclusion The CoxA5VP4 region of pathogen of HFMD in Xicheng District, Beijing was closely related to Chinese HQ728261 in 2009 with no significant variation in its sequence.