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目的研究2015年山东省烟台地区手足口病(hand,foot,and mouth disease,HFMD)的病原谱,并分析病原谱中柯萨奇病毒A组16型(Coxsackievirus A16,CV-A16)流行株的进化及VP1区重要氨基酸位点的变异情况。方法采用实时荧光逆转录聚合酶链反应方法,对2015年烟台地区采集于HFMD患者的696份样品进行肠道病毒(enterovirus,EV)核酸检测和分子定型。根据EV的优势血清型,选取10株CV-A16扩增VP1区,并进行核苷酸序列及系统发育分析。结果从696份标本中检出EV 412株,其中101株为CV-A16,76株为EV-A71型。10株CV-A16的核苷酸序列相似性为88.4%~99.9%,氨基酸序列相似性为98.6%~100%。10株CV-A16烟台株分别属于B基因型的B1a和B1b进化分支,其中B1b进化分支为优势株。与参考株Tainan/5079/98(AF177911)相比,10株CVA16分离株氨基酸序列的第11、23、99、145、289位氨基酸出现突变,10株分离株的这些位点也有不同,表明烟台地区的CV-A16有其独特的突变位点。结论 CV-A16是引起2015年烟台地区HFMD发病的常见病原体,本次分离到的CV-A16均属于B基因型的Bla和B1b进化分支。
Objective To study the pathogen spectrum of hand, foot and mouth disease (HFMD) in Yantai, Shandong Province in 2015 and to analyze the prevalence of Coxsackievirus A16 (CV-A16) Evolution and Variations of Important Amino Acids in VP1 Region. Methods Real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR) was used to detect and molecularize the enterovirus (EV) nucleic acid in 696 samples from patients in HFMD in Yantai in 2015. According to the predominant serotype of EV, 10 strains of CV-A16 were selected to amplify the VP1 region, and nucleotide sequence and phylogenetic analysis were performed. Results EV 412 strains were detected from 696 specimens, of which 101 were CV-A16 and 76 were EV-A71. The nucleotide sequence identities of 10 CV-A16 strains ranged from 88.4% to 99.9% with similar amino acid sequence of 98.6% -100%. Ten CV-A16 Yantai strains belonged to the B1a and B1b clades of B genotype respectively, of which B1b was the dominant one. Compared with the reference strain Tainan / 5079/98 (AF177911), amino acids 11, 23, 99, 145 and 28 of the 10 CVA16 isolates were mutated, and the ten isolates also showed different positions, indicating that Yantai The region’s CV-A16 has its own unique mutation site. Conclusions CV-A16 is a common causative agent of HFMD in Yantai in 2015. The isolated CV-A16 belongs to the Bla and B1b clades of B genotype.