2018—2020年黔东南少数民族自治州肠道病毒A71型基因特征分析  

Genetic characteristics of enterovirus A71 in Qiandongnan Miao and Dong Autonomous Prefecture from 2018 to 2020

作　　者：肖俊[1] 单竹周 江忠晶 裴峰锋 王宇 唐小敏[3] XIAO Jun;SHAN Zhuzhou;JIANG Zhongjing;PEI Fengfeng;WANG Yu;TANG Xiaomin(Qiandongnan Center for Disease Control and Prevention,Qiandongnan,Guizhou 556000,China)

机构地区：[1]黔东南州疾病预防控制中心,贵州556000 [2]贵阳市第一人民医院检验科 [3]贵州省疾病预防控制中心

出　　处：《应用预防医学》2023年第4期222-226,231,共6页Applied Preventive Medicine

基　　金：国家自然科学基金项目(81860594);贵州省科技计划项目(黔科合基础[2018]1094);筑科合同[2021]43-25号。

摘　　要：目的对2018—2020年黔东南州手足口病(hand,foot and mouth disease,HFMD)进行鉴定分析,并对肠道病毒A组71型(enterovirus A71,EV-A71)VP1编码区进行基因特征分析。方法2018—2020年黔东南州共采集HFMD病例标本2789份,首先,采用荧光定量RT-PCR初筛肠道病毒核酸,阳性病例标本采用人类横纹肌肉瘤(human rhabdomyosarcoma,RD)细胞进行病毒分离,对分离到的病毒株提取病毒核酸,经RT-PCR方法扩增VP1编码区核苷酸序列并进行序列测定。使用MEGA 11.0软件构建遗传进化树进行基因特征分析。结果2789份HFMD标本中检出EV-A71核酸阳性标本为328份,阳性率为11.8%。从部分EV-71阳性患者的咽拭子及肛拭子中,分离培养获得7株病毒株,其中6株来自于轻症病例,另1株为重症病例,均属于EV-A71基因型C4a基因亚型,分成2个不同的簇。7株EV-A71 VP1序列之间核苷酸和氨基酸同源性分别为89.8%~99.9%和98.0%~99.7%;与BrCr原型参考株间的核苷酸和氨基酸同源性分别为77.1%~78.8%和94.8%~96.6%;并在VP1有18处氨基酸位点发生变异。结论2018—2020年黔东南州EV-A71分离株属于C4a基因亚型,多处氨基酸位点存在变异,可导致多条传播链循环。Objective To identify and analyze the hand,foot,and mouth disease(HFMD)in Qiandongnan Prefecture from 2018 to 2020,and analyze the genetic characteristics of VP1 coding region of enterovirus A71(EV-A71).Methods A total of 2789 HFMD case specimens were collected in Qiandongnan Prefecture from 2018 to 2020.First,fluorescent quantitative RT-PCR was used to screen the enterovirus nucleic acid.The positive case samples were isolated from human rhabdomyosarcoma(RD)cells,and the virus nucleic acid was extracted from the isolated virus strain.The nucleotide sequence of VP1 coding region was amplified by RT-PCR and sequenced.Genetic phylogenetic trees were constructed by MEGA 11.0 software for gene feature analysis.Results Among the 2789 HFMD samples,328 were positive for EVA71 nucleic acid,with a 11.8%positivity rate.From throat swabs and anal swabs of some EV-71 positive patients,7 virus strains were isolated and cultured,and 6 of them were mild cases and one was severe case,all of which belonged to the EV-A71 genotype C4a gene subtype,divided into 2 distinct clusters.The nucleotide and amino acid homology among the 7 strains of EV-A71 VP1 sequences were 89.8%-99.9%and 98.0%-99.7%,respectively.The nucleotide and amino acid homology with the BrCr prototype reference strain were 77.1%-78.8%and 94.8%-96.6%,respectively.In addition,there were 18 amino acid positions in VP1 that mutated.Conclusions From 2018 to 2020,the EV-A71 isolates in Qiandongnan Prefecture belonged to the C4a gene subtype,and there were mutations in multiple amino acid positions,which could lead to the circulation of multiple transmission chain.

关 键 词：肠道病毒 肠道病毒A组71型(EV-A71) 基因型 手足口病(HFMD) 

分 类 号：R373.2[医药卫生—病原生物学]