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目的了解2012年北京市某幼儿园手足口病暴发疫情的病原分子特征。方法采集病例咽拭子标本,利用RD细胞和Hep-2细胞进行病毒分离。RT-PCR扩增肠道病毒VP1区基因序列,生物信息学方法鉴定基因型别并进行种系发生分析。结果 13例患者样本中有11件为非肠道病毒71型(EV71)非柯萨奇病毒A16型(CVA16)肠道病毒阳性,其中9例患者咽拭子鉴定为柯萨奇病毒A10型(CVA10),2例样本基因分型失败。9份咽拭子样本分离出7株CVA10毒株。VP1区基因序列分析表明:本起疫情分离株核苷酸和氨基酸的同源性分别为99.8%～100%,99.7%～100%。与2009年山东CVA10分离株VP1区核苷酸和氨基酸的同源性分别为94.8%～96.3%,98.3%～99.3%,与2010年法国CVA10分离株及CVA10原型株(Kowalik株)VP1区核苷酸和氨基酸同源性分别为72.6%～78.1%,91.1%～93.3%。结论本起手足口暴发疫情由CVA10毒株引起。该毒株与山东2009年CVA10分离株亲缘关系较近。
Objective To understand the molecular characteristics of pathogens of HFMD outbreak in a nursery in Beijing in 2012. Methods Throat swab specimens were collected, and RD cells and Hep-2 cells were used for virus isolation. The sequence of VP1 gene of enterovirus was amplified by RT-PCR, and the genotypes were identified by bioinformatics method and phylogenetic analysis was performed. Results Eleven of the 13 patient samples were positive for non-enterovirus 71 (EV71) non-Coxsackie A16 (CVA16) enterovirus, of which 9 were throat swabs identified as Coxsackie A10 CVA10), 2 samples failed to genotype. Seven CVA10 strains were isolated from 9 throat swab samples. Sequence analysis of VP1 gene showed that the nucleotide and amino acid homology of the isolates were 99.8% -100% and 99.7% -100%, respectively. The nucleotide and amino acid identities of VP1 of CVA10 isolates from Shandong province in 2009 were 94.8% -96.3% and 98.3% -99.3%, respectively. The homology with VP1 region of CVA10 strain and Kowalik strain in 2010 The nucleotide and amino acid homologies were 72.6% -78.1% and 91.1% -93.3%, respectively. Conclusion The onset of foot-mouth outbreak was caused by CVA10 strain. The strain and Shandong CVA10 2009 isolates closer genetic relationship.