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目的分析云南省2012年引起手足口病(hand-foot-and-mouth disease,HFMD)流行的柯萨奇病毒A组16型(coxsackievirus A16,CV-A16)的基因特征,为今后开展CV-A16的分子流行病学研究和疾病防控提供数据。方法对从云南省2012年HFMD患者样本中分离到的CV-A16,采用逆转录-聚合酶链反应扩增VP1基因片段全长,并对扩增产物进行测序。根据VP1基因核苷酸序列结果,从中选取20株与其他CV-A16毒株序列(来源于基因库)构建亲缘性关系树,并且进行核苷酸和氨基酸同源性分析。结果从云南省2012年HFMD患者样本中共分离到112株CV-A16,均为B1基因型,其中102株为B1b亚型,10株为B1a亚型,选取所有的B1a和10株B1b进行分析,核苷酸和氨基酸的同源性分别为89.10%~97.87%和98.91%~100.00%。结论引起2012年云南省HFMD流行的CV-A16毒株属于B1基因型的两大进化分支B1a和B1b,两个进化分支在云南共同进化和流行。
Objective To analyze the genetic characteristics of coxsackievirus A16 (CV-A16) in Yunnan province in 2012 caused by hand-foot-and-mouth disease (HFMD) Molecular epidemiological studies and disease control provide data. Methods The total length of VP1 gene fragment was amplified by RT-PCR from CV-A16 isolated from samples of HFMD patients in Yunnan province in 2012, and the amplified products were sequenced. According to the results of nucleotide sequence of VP1, 20 phylogenetic trees were constructed from the sequences of other CV-A16 strains (derived from gene bank), and nucleotide and amino acid homology analysis was performed. Results A total of 112 CV-A16 strains were isolated from samples of HFMD patients in Yunnan Province in 2012. All of them were B1 genotypes, of which 102 were B1b subtypes and 10 were B1a subtypes. All B1a and 10 B1b strains were selected for analysis. The nucleotide and amino acid homologies were 89.10% -97.87% and 98.91% -100.00%, respectively. Conclusion The CV-A16 strains that caused HFMD epidemic in Yunnan Province in 2012 belonged to B1a and B1b of B1 genotype, and the two evolutionary clades evolved and popularized in Yunnan.