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目的分析2013年珠海市手足口病患儿柯萨奇病毒A2(CV-A2)5′UTR-VP4-VP2区基因特征,探讨其与国内外其他地区CV-A2分离株的关系。方法采集2013年珠海市手足口病患儿粪便标本,提取病毒RNA,用半巢式RT-PCR方法扩增5′UTR-VP4-VP2区,通过测序和Blast比对确定病毒型别,并利用DNAStar软件和MEGA4.1软件分别对珠海市CV-A2分离株进行核苷酸序列分析和遗传进化树分析。结果共鉴定出4株CV-A2,核苷酸序列分析表明CV-A2珠海株与2012年中国香港株在5′UTR-VP4-VP2区有最高的同源性,达97.4%~98.2%;进化树分析显示4株CV-A2珠海株与2012年中国香港株共同构成A进化树分支,Bootstrap值为99%。结论珠海市4株CV-A2与2012年中国香港株在5′UTR-VP4-VP2区具有较高的同源性,且进化关系密切。
Objective To analyze the characteristics of 5 ’UTR-VP4-VP2 gene of coxsackievirus A2 (CV-A2) in children with hand-foot-mouth disease in Zhuhai in 2013 and its relationship with CV-A2 isolates from other regions of China. Methods Stool samples of hand, foot and mouth disease were collected from Zhuhai City in 2013. The viral RNA was extracted and the 5’UTR-VP4-VP2 region was amplified by semi-nested RT-PCR. The virus type was determined by sequencing and Blast comparison. DNAStar software and MEGA4.1 software were used for nucleotide sequence analysis and genetic tree analysis of Zhuhai CV-A2 isolates respectively. Results A total of 4 strains of CV-A2 were identified. Nucleotide sequence analysis showed that the highest homology between CV-A2 Zhuhai strain and the Hong Kong strain in China in 2012 was 97.4% -98.2% in the 5’UTR-VP4-VP2 region. Phylogenetic tree analysis showed that four strains of CV-A2 Zhuhai and Hong Kong, China in 2012 formed an A-phylogenetic tree with a Bootstrap value of 99%. Conclusion The four strains of CV-A2 in Zhuhai have a high homology with the Hong Kong strain of China in the region of 5’UTR-VP4-VP2 in 2012, which is closely related to evolution.