云南省3株柯萨奇A组16型病毒全基因组序列分析  

Analysis of whole genome sequence of three coxsackievirus A16strains isolated from Yunnan Province,China

作　　者：冯昌增[1] 刘昕蓓 张名[1] 许丹菡 徐炽 何云龙 马绍辉[1] FENG Chang-zeng;LIU Xin-bei;ZHANG Ming;XU Dan-han;XU Chi;HE Yun-long;MA Shaohui(Key Laboratory for Research and Development of Major Infectious Diseases Vaccine in Yunnan,Institute of Medical Biology,Chinese Academy of Medical Sciences,Peking Union Medical College,Yunnan Kunming,650118,China;School of Medicine,Yunnan University)

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]云南大学医学院

出　　处：《中国病原生物学杂志》2022年第2期149-153,158,共6页Journal of Pathogen Biology

基　　金：云南省科技厅科技计划项目(No.202002AA100009)。

摘　　要：目的分析柯萨奇病毒A组16型(coxsackievirus A16,CV-A16)分离株全基因组特征。方法使用人横纹肌肉瘤细胞(human rhabdomyosarcoma,RD)、非洲绿猴肾细胞(Vero)和人胚肺二倍体细胞(human embryonic lung diploid fibroblasts,KMB17)从2019年云南手足口病患者粪便标本中分离CV-A16毒株。采用RT-PCR扩增其全基因组,测序并通过生物信息学方法对其全基因序列、系统进化和基因重组进行分析。结果共分离到3株CV-A16毒株,均为Vero细胞分离株。VP1系统进化分析表明,K106/YN/CHN/2019和K39/YN/CHN/2019属于B1基因亚型的B1b分支,K23/YN/CHN/2019属于B1基因亚型的B1a分支。在全基因组核苷酸序列上,K106/YN/CHN/2019和K39/YN/CHN/2019与中国CV-A16分离株相似性高,K23/YN/CHN/2019与国外CV-A16分离株相似性较高,其中,与澳大利亚分离株C138/CHW/AUS/2016之间的核苷酸相似性为97.6%。基于P1、P2和P3的系统进化、全基因组同源性和重组分析结果显示CV-A16在5'-UTR及P2、P3区可能与肠道病毒A组多个其他血清型病毒发生过型间重组事件。结论2019年从云南省手足口病患者粪便标本中分离到3株CV-A16,均为中国大陆流行基因型,为CV-A16的分子流行病学研究提供了理论基础。Objective To isolate Coxsackievirus A16(CV-A16)strains from fecal samples of patients with hand-footmouth disease(HFMD)in Yunnan in 2019,and analyze their whole genome sequences and phylogenetic characteristics.Methods Three cell lines,human rhabdomyosarcoma(RD),green monkey kidney(Vero)and human embryonic lung diploid fibroblasts(KMB17),were used for viral isolation.The whole genome sequences were amplified by reverse transcription-polymerase chain reaction(RT-PCR)and sequenced using the Sanger sequencing method.The similarities of nucleotide and amino acid sequences were calculated using Geneious Prime 2020.1.2.Phylogenetic analysis was performed using the Neighbor-Joining method in MEGA 7.0.Simplot 3.5.1was used to carry out SimPlot and Bootscan analyses.Results In the present study,3novel CV-A16strains were isolated using Vero cells.The phylogenetic analysis based on the VP1sequence showed that two of the strains(K106/YN/CHN/2019and K39/YN/CHN/2019)belonged to B1bclade of subgenotype B1,while the K23/YN/CHN/2019strain belonged to B1aclade of subgenotype B1.In whole genome,K106/YN/CHN/2019and K39/YN/CHN/2019shared high nucleotide similarities with the CV-A16strains isolated from China.But,K23/YN/CHN/2019had high nucleotide similarities with CV-A16isolates outside of China,especially the highest identities with Australia strain C138/CHW/AUS/2016(97.6%).The phylogenetic analysis based on the P1,P2and P3,whole genome homology analysis and recombination analysis suggested that inter-serotypic recombination events may have occurred between the CV-A16and some other serotypes of Enterovirus A in 5'-UTR P2and P3.Conclusion In this study,3novel CV-A16strains were isolated from stool samples of HFMD patients in Yunnan Province,all of which belonged to the dominant genotypes in China's Mainland.The results of the analysis of the whole genome sequence and phylo-genetic characteristics may provide laboratory data for further investigation of the prevalence,transmission and molecular evolution of CV-A16.

关 键 词：柯萨奇病毒A组16型 全基因组序列 型间重组 

分 类 号：R383.2[医药卫生—医学寄生虫学]