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目的了解福州地区流行的柯萨奇病毒A10型(CV-A10)的VP1区基因特征及种系进化分布。方法采集临床诊断为手足口病患者的咽拭子或肛拭子样本,通过real-time PCR法确定样本的肠道病毒型别。选取部分CV-A10的临床样本,利用RD细胞进行病毒分离,RT-PCR扩增肠道病毒的VP1基因片段,并进行序列测定,利用DNAStar和MEGA 5.1软件进行核苷酸和氨基酸同源性分析并构建亲缘进化树。结果检测出的1 951份肠道病毒阳性样本中,肠道病毒71型(EV-A71)、柯萨奇病毒A16型(CVA16)、CV-A10的阳性率分别为24.1%,16.7%,9.4%,不同肠道病毒阳性率差异有统计学意义。福州市CV-A10型各分离株的核苷酸和氨基酸同源性分别为94.0%~99.9%和97.7%~100.0%,与CV-A10原型株(Kowalik株)的同源性分别为76.4%~76.7%和91.9%~93.3%。亲缘进化树的分析结果显示福州地区CV-A10分离株属于D亚型。结论 CV-A10是引起福州地区手足口病的常见病原体之一,本次分离到的CV-A10病毒株均属于D基因型。
Objective To understand the characteristics and phylogenetic distribution of VP1 gene of Coxsackievirus A10 (CV-A10) in Fuzhou. Methods Samples of throat swabs or anal swabs clinically diagnosed as hand-foot-mouth disease were collected, and the enterovirus types of samples were determined by real-time PCR. The clinical samples of partial CV-A10 were selected and the virus was isolated by RD cells. The VP1 gene fragment of enterovirus was amplified by RT-PCR and sequenced. The DNAStar and MEGA 5.1 software were used to analyze the nucleotide and amino acid homology And build phylogenetic tree. Results The positive rates of enterovirus 71 (EV-A71), Coxsackie virus A16 (CVA16) and CV-A10 were 24.1%, 16.7%, 9.4% %, The positive rate of different enterovirus differences were statistically significant. The nucleotide and amino acid homologies of CV-A10 isolates in Fuzhou were 94.0% -99.9% and 97.7% -100.0%, respectively, and those of CV-A10 isolates (Kowalik strain) were 76.4% ~ 76.7% and 91.9% ~ 93.3%. Phylogenetic tree analysis showed that the CV-A10 isolates in Fuzhou belonged to D subtype. Conclusions CV-A10 is one of the common pathogens causing hand-foot-mouth disease in Fuzhou. The CV-A10 strains isolated in this study all belong to D genotype.