摘要
目的 分析2016-2022年银川市手足口病(HFMD)肠道病毒的病原构成,了解主要流行株柯萨奇病毒(CV)A组4型(CV-A4)、6型(CV-A6)、10型(CV-A10)和16型(CV-A16)分子病原学进化特征.方法 收集2016-2022年银川市HFMD监测病例标本,采用RT-qPCR鉴定肠道病毒型别,病毒分离获得的毒株采用RT-PCR扩增其VP1区,通过测序和比对进行序列分析,使用DNASTAR Lasergene软件进行序列拼接整理,用MEGA 7.0软件构建系统进化树.结果 2016-2022年共检测手足口病例标本2 492份,检出肠道病毒核酸阳性标本1 923份,总阳性率为77.2％.共检出肠道病毒型别12种,不同年份病原分布差异有统计学意义(P＜0.001).银川市 CV-A4、CV-A6、CV-A10 和 CV-A16 分离株分别属于 C2(Cluster Ⅲ分支)、D3a、C、B1a/B1b基因组.与原型株VP1区序列比较,分别出现了 8、32、24和26个氨基酸位点的变异.结论 2016-2022年银川市HFMD病原存在多种肠道病毒共流行的模式,以EV-A71、CV-A16和CV-A6为优势病原动态变化,交替流行.应使用更加全面的肠道病毒基因分型方法持续监测病原谱的改变.
Abstract
Objective To analyze the pathogenic composition and characteristics of molecular etiology of coxsackievirus A4(CV-A4),coxsackievirus A6(CV-A6),coxsackievirus A10(CV-A10),and coxsackievirus A16(CV-A16)of enterovirus in cases of hand,foot and mouth disease(HFMD)in Yinchuan between 2016 and 2022.Methods The specimens were collected from HFMD surveillance cases in Yinchuan from 2016 to 2022.The enterovirus genotypes were determined by RT-qPCR assay.The VP1 regions of virus isolating strains were amplified by RT-PCR and sequenced.Sequences splicing were performed using biological software DNASTAR lasergene and the phylogenetic trees were constructed by software MEGA 7.0.Results A total of 2 492 HFMD cases were tested during the specified time period,resulting in the detection of 1 923 enterovirus nucleic acid positive samples,yielding an overall positive rate of 77.2％.Furthermore,12 different types of enterovirus were identified,and a significant difference in the distribution of enterovirus types by year was observed(P＜0.001).In Yinchuan,the CV-A4,CV-A6,CV-A10,and CV-A16 isolates were found to belong to the C2(Cluster Ⅲbranch),D3a,C,and B1a/B1b gene clusters,respectively.A comparison of their VP1 region sequences with the prototype revealed the presence of 8,32,24 and 26 amino acid site mutations,respectively.Conclusion Between 2016 and 2022,aco-circulation pattern of multiple enteroviruses was observed in HFMD cases in Yinchuan,with EV-A71,CV-A16,and CV-A6 being the dominant enteroviruses that underwent dynamic changes and alternate spread.Therefore,more comprehensive methods for genotyping enterovirus should be used to continuously monitor changes in the pathogenic spectrum.
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