2019年云南省昆明市柯萨奇病毒A组4型毒株分离鉴定及其VP1基因特性分析  

Isolation,identification and characterization of VP1 gene of Coxsackievirus A4 in Kunming City,Yunnan Province,China in 2019

作　　者：丛山日 刘红波 许丹菡 张名[1] 冯昌增[1] 黄小琴[1] 孙浩[1] 杨昭庆[1] 马绍辉[1] CONG Shan-ri;LIU Hong-bo;XU Dan-han;ZHANG ming;FENG Chang-zeng;HUANG Xiao-qin;SUN Hao;YANG Zhao-qing;MA Shao-hui(Key Laboratory for Research and Development of Major Infectious Diseases Vaccine in Yunnan,Institute of Medical Biology,Chinese Academy of Medical Sciences and Peking Union Medical College,Kunming 650118,Yunnan Province,China)

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室,云南昆明650118 [2]威海市人力资源和社会保障局,山东威海5323368

出　　处：《中国生物制品学杂志》2022年第2期144-149,158,共7页Chinese Journal of Biologicals

基　　金：云南省科技计划项目(2017FA006).

摘　　要：目的对2019年云南省手足口病(hand,foot and mouth disease,HFMD)患者粪便中分离获得的柯萨奇病毒A组4型(coxsackievirus A4,CV-A4)毒株的VP1基因遗传特征进行分析。方法利用人横纹肌肉瘤RD细胞、人胚肺二倍体KMB-17细胞和非洲绿猴肾Vero细胞分离病毒,以病毒RNA作为逆转录-聚合酶链反应(reverse transcriptionpolymerase chain reaction,RT-PCR)的模板,扩增病毒完整VP1基因,利用MEGA 6.06和Geneious 9.0.2等软件对VP1序列进行分析。结果选取8株CV-A4分离株进行分析,其完整VP1序列为915 bp,编码305个氨基酸。8株CV-A4云南分离株与CVA4/High Point/USA/1949/AF081295(简称原型株)的核苷酸一致性和氨基酸同源性分别为83.4%~84.6%和97.0%~98.0%;与其他国内CV-A4参考株相比,核苷酸一致性和氨基酸同源性分别为87.7%~97.0%和97.7%~99.3%。8株CV-A4云南分离株相互比对,核苷酸一致性和氨基酸同源性分别为91.0%~99.9%和97.7%~100.0%。CV-A4云南分离株与大部分中国分离株同属C2基因亚型。结论本研究中CV-A4分离株均是中国大陆地区优势流行株。Objective To analyze the genetic characteristics of VP1 gene of Coxsackievirus A4(CV-A4)strains isolated from the feces of patients with hand,foot and mouth disease(HFMD)in Kunming City,Yunnan Province,China in 2019.Methods Virus was isolated with RD,KMB-17 and Vero cells,from which the viral RNA was extracted and used as the template for amplification of whole VP1 gene by RT-PCR.MEGA 6.06 and Geneious 9.0.2 software were used to analyze the whole VP1 gene sequence.Results Eight CV-A4 isolates were selected,of which the whole VP1 sequence consisted of 915 bp encoding 305 amino acids.The homologies of nucleotides and deduced amino acids of the eight isolates were 83.4%~84.6%and 97.0%~98.0%to those of CVA4/High Point/USA/1949/AF081295(prototype),and 87.7%~97.0%and 97.7%~99.3%to those of other domestic CV-A4 reference strains,respectively.However,the homologies of nucleotides and deduced amino acids of the eight isolates were 91.0%~99.9%and 97.7%~100.0%respectively.Like most of Chinese isolates,the eight isolates belonged to C2 gene subtype.Conclusion The CV-A4 isolates were the predominant epidemic strains in China.

关 键 词：柯萨奇病毒A组4型 分离 鉴定 VP1序列 进化分析 遗传特性 

分 类 号：R373.[医药卫生—病原生物学]