2018－2019年安徽省柯萨奇病毒A组16型B1基因亚型分离株分子特征分析

史永林; 葛盈露; 马婉婉; 孙永; 吴家兵; 苏斌

doi:10.3784/jbjc.202203140097

2018－2019年安徽省柯萨奇病毒A组16型B1基因亚型分离株分子特征分析

Molecular characterization of coxsackievirus A16 B1 in Anhui, 2018−2019

摘要

摘要:
目的了解安徽省2018—2019年手足口病（HFMD）柯萨奇病毒A组16型（coxsackievirus A16，CV-A16）分子病原学进化特征。
方法对安徽地区2018—2019年从HFMD患者咽拭子样本中分离的CV-A16毒株，采用反转录聚合酶链式反应（RT-PCR）扩增其VP1基因全长核酸片段、测定其核苷酸序列。采用肠道病毒在线分型工具鉴定CV-A16毒株基因亚型，运用MEGA 6.0生物软件最大似然法（maximum likelihood method）构建 VP1基因进化树、MegAlign比较核苷酸和氨基酸同源性。
结果安徽省2018—2019年共分离到HFMD CV-A16毒株88株。其中2018年分离到30株，2019年58株。其VP1区全长基因均由891个核苷酸碱基组成，编码297个氨基酸。基因分型鉴定和进化分析显示，88株 CV-A16毒株中B1a和B1b基因亚型分别为25株（28.41%）、63株（71.59%）。 VP1区遗传进化表明，88株CV-A16毒株均归为B1基因型，其中 25株属于B1a基因亚型，63株属于B1b基因亚型。同源性分析显示，B1a基因亚型毒株与CV-A16原型株G-10核苷酸同源性为64.1%～65.3%，氨基酸同源性为90.3%～91.1%； B1b毒株与原型株核苷酸及氨基酸同源性分别为62.8%～66.0%和91.1%～91.8%。 88株CV-A16与原型株VP1区氨基酸序列比较发现26个氨基酸位点出现变异。
结论安徽地区2018—2019年HFMD CV-A16毒株以B1b与B1a共同流行，B1b为优势基因亚型，应重视CV-A16流行毒株的分子病原学监测。

Abstract:
Objective To understand the characteristics of molecular etiology of coxsackievirus A16 (CV-A16) causing hand foot and mouth disease (HFMD) in Anhui province from 2018 to 2019.
Methods CV-A16 strains were isolated from throat swabs of HFMD cases in Anhui during 2018−2019. The full-length gene of VP1 of CV-A16 strains was amplified by RT-PCR and sequenced. The genotype/sub-genotype of CV-A16 strains was identified by enterovirus online typing tool (https://www.rivm.nl/mpf/typingtool/enterovirus/), and the phylogenetic tree of VP1 gene was constructed by maximum likelihood method with biological software MEGA 6.0 Software Megalign was used to compare the homology of nucleotide and amino acid.
Results A total of 88 CV-A16 strains were isolated from HFMD cases in Anhui from 2018 to 2019. Among the strains, 30 were isolated in 2018 and 58 were isolated in 2019. The full-length gene of VP1 regions of the strains were all composed of 891 nucleoside acid, encoding 297 amino acids. Genotyping identification and phylogenetic analysis showed that among the 88 CV-A16 strains, B1a and B1b sub-genotypes were detected in 25 and 63 strains, accounting for 28.41% (25/88) and 71.59% (63/88) respectively. The genetic evolution of VP1 region showed that 88 CV-A16 strains were classified as B1 genotype, in which 25 belonged to sub-genotype B1a and 63 belonged to sub-genotype B1b. Homology analysis showed that the nucleotide homology between B1a strain and CV-A16 prototype strain G-10 was 64.1%–65.3%, and the amino acid homology was 90.3%–91.1%; The homology of nucleotide and amino acid between B1b strain and prototype strain was 62.8%–66.0% and 91.1%–91.8%, respectively. The amino acid sequences of VP1 regions of 88 strains CV-A16 were compared with that of the prototype strain, and mutation occurred in 26 amino acid loci.
Conclusion B1b and B1a were the common sub-genotypes of CV-A16 causing HFMD in Anhui from 2018 to 2019, and B1b was predominant. It is necessary to pay attention to the surveillance for molecular etiology of CV-A16 epidemic strains.

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