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目的对青海省2013年手足口病(Hand,Foot and Mouth Disease;HFMD)的病原谱和人肠道病毒A组71型(Human Enterovirus Group A Type 71,HEV_(A71))的基因特征进行分析。方法对青海省2013年送检的HFMD标本,用荧光定量逆转录-聚合酶链反应(Reverse Transcription-Polymerase Chain Reaction,RT-PCR)方法进行筛查,对HEV_(A71)阳性标本进行病毒分离,对分离到的病毒提取核酸,用RT-PCR方法对VP1编码区进行扩增及核苷酸序列测定和整理分析,并与HEV_(A71)各基因型和基因亚型的代表株序列构建亲缘关系进化树。结果对青海省2013年分离的62株HEV_(A71)进行VP1编码区核苷酸序列测定后,分析发现它们在VP1编码区核苷酸水平和氨基酸水平的同源性分别为95.5%~100.0%和99.0%~100.0%。与其他各基因型和基因亚型的HEV_(A71)代表株构建的亲缘进化树中显示,青海省HEV_(A71)分离株与C4亚型代表株聚为一簇,属于C4基因亚型C_(4a)进化分支。与2008年、2012年在青海省分离的HEV_(A71)在核苷酸和氨基酸水平的同源性分别为95.8%~97.5%和99.0%~99.7%。结论2013年,C_(4a)亚型HEV_(A71)在青海省持续传播并占有绝对优势,且存在两条传播链的流行。
Objective To analyze the pathogenic profiles of Hand, Foot and Mouth Disease (HFMD) in Qinghai Province in 2013 and the gene characteristics of Human Enterovirus Group A Type 71 (HEV_ (A71)). Methods HFMD specimens from 2013 in Qinghai Province were screened by RT-PCR and HEV_ (A71) positive samples were isolated by RT-PCR. Nucleic acid was extracted from the isolated virus, and the VP1 coding region was amplified by RT-PCR and the nucleotide sequence was determined and analyzed. The genetic relationship was also established with the representative strains of HEV_ (A71) genotypes and subtypes Evolutionary tree. Results The nucleotide sequences of the VP1 coding region of 62 HEV_ (A71) isolates isolated in 2013 in Qinghai Province were analyzed. The homologies of nucleotide and amino acid sequences in the VP1 coding region were 95.5% -100.0% And 99.0% to 100.0%. The phylogenetic tree constructed with the HEV_ (A71) representative strains of other genotypes and subgenotypes showed that the HEV_ (A71) isolate and the C4 subtype of Qinghai province clustered into clusters, belonging to the C4 subtype C_ ( 4a) Evolutionary branch. The nucleotide and amino acid homology of HEV_ (A71) isolated from Qinghai province in 2008 and 2012 were 95.8% -97.5% and 99.0% -99.7%, respectively. Conclusions In 2013, the C_ (4a) subtype HEV_ (A71) continued to spread and occupy an absolute advantage in Qinghai Province, and there was a prevalence of two transmission chains.