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目的对2009年中国云南昆明地区柯萨奇病毒A组16型(Coxsackievirus A16,CoxA16)毒株的部分VP1区进行分析,了解CoxA16的遗传特性。方法设计针对肠道病毒A群的通用引物,采用半巢式RT-PCR扩增目的片段,对PCR产物测序;使用Omiga和Mega4.1等生物学软件进行核苷酸、氨基酸序列比对及病毒的系统进化分析。结果200份2009年昆明市儿童医院临床诊断为手足口病或并发脑炎疑似肠道病毒感染的患者粪便标本中有52份为CoxA16感染,其中15份来自脑炎患者。52株CoxA16病毒部分VP1区核苷酸和氨基酸序列同源性均较高,核苷酸序列同源性为98.5%～100%,氨基酸序列同源性为98.8%～100%;与shzh00-2、shzh99-48和107/Toyama/1990相比较,核苷酸序列同源性为92.3%～93.8%,氨基酸序列同源性为95.3%～96.8%;与国际标准株G10相比较,核苷酸序列同源性为78.5%～80.3%,氨基酸序列同源性为93.2%～94.5%;与其他13个参考株的核苷酸序列同源性为96.6%～99.5%,氨基酸序列同源性为98.3%～100%。基因系统进化分析显示,52株CoxA16均属于B基因型的一个分支。结论 2009年中国云南昆明地区CoxA16株属于B2亚型。
Objective To analyze the partial VP1 region of Coxsackievirus A16 (CoxA16) strain in Kunming, Yunnan Province, China in 2009 to understand the genetic characteristics of CoxA16. Methods A universal primer designed for Enterovirus A group was designed. The target fragment was amplified by semi-nested RT-PCR and the PCR products were sequenced. Nucleotide and amino acid sequence alignment and biological analysis were performed using biological software such as Omiga and Mega4.1 System evolution analysis. RESULTS: Of the 200 stool samples from 200 stool samples clinically diagnosed as hand, foot and mouth disease or suspected enterovirus infection in Kunming Children’s Hospital in 2009, 52 were CoxA16 infection, 15 of which were from encephalitis patients. The homology of nucleotide and amino acid sequences of VP1 of 52 CoxA16 viruses was high, with 98.5% -100% nucleotide sequence homology and 98.8% -100% amino acid sequence homology with that of shzh00-2 , Shzh99-48 and 107 / Toyama / 1990, the nucleotide sequence homology was 92.3% -93.8%, and the amino acid sequence homology was 95.3% -96.8%. Compared with the international standard strain G10, the nucleotide The sequence homology was 78.5% -80.3%, and the amino acid sequence homology was 93.2% -94.5%. The nucleotide sequence homology with other 13 reference strains was 96.6% -99.5%, and the amino acid sequence homology was 98.3% ~ 100%. Phylogenetic analysis showed that 52 CoxA16 belonged to a branch of B genotype. Conclusion In 2009, CoxA16 strain in Kunming, Yunnan Province of China belonged to B2 subtype.