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目的了解恩施和武汉地区手足口病(HFMD)的分子流行病学特征和病原体柯萨奇病毒A16型(CA16)VP1的基因特征。方法收集2010年4月至2011年8月来自于恩施和武汉地区的189例HFMD患儿咽拭子标本,接种病毒易感Vero细胞,对产生明显细胞病变效应的标本进行初筛和PCR鉴定;对阳性毒株VP1基因序列进行扩增,构建pMD18-T-VP1重组克隆,然后测定其VP1全长序列,通过MEGA5软件构建系统进化树进行系统进化分析,通过DNAstar软件对VP1保守表位的氨基酸序列保守性进行分析。结果手足口病患者以≤5岁婴幼儿为主,男性多于女性。从189例手足口病患儿咽拭子标本中分离到7株CA16,VP1序列进化分析显示均属于B1亚型。对VP1基因上的6个抗原表位进行分析,其中的3个区域呈现高度保守性,这些保守区可被用于疫苗研究。结论 CA16是2010-2011年恩施和武汉地区手足口病的重要病原体之一,属于B1基因型。
Objective To investigate the molecular epidemiological characteristics of HFMD in Enshi and Wuhan and the genetic characteristics of the pathogen Coxsackie virus A16 (CA16) VP1. Methods Throat swab samples of 189 children with HFMD from Enshi and Wuhan were collected from April 2010 to August 2011, and the virus susceptible Vero cells were inoculated and the specimens with obvious cytopathic effect were screened and identified by PCR. The VP1 gene sequence of the positive strain was amplified to construct the recombinant clone pMD18-T-VP1, and then the full-length VP1 sequence was determined. Phylogenetic tree was constructed by using MEGA5 software. The amino acid sequence of the VP1 conserved epitope Sequence conservation analysis. Results Hand, foot and mouth disease patients were less than 5 years old infants and young children, more men than women. Seven strains of CA16 were isolated from throat swab samples of 189 children with hand-foot-mouth disease. The evolutionary analysis of VP1 sequences showed that all belong to B1 subtype. Three antigenic epitopes on the VP1 gene were analyzed. Three of these epitopes were highly conserved and these conserved regions could be used for vaccine research. Conclusion CA16 is one of the important pathogens of hand-foot-mouth disease in Enshi and Wuhan in 2010-2011 and belongs to B1 genotype.