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16型(Coxsackievirus A16,CVA16)毒株的全VP1基因遗传特征进行分析.方法 使用人胚肺二倍体KMB-17细胞和非洲绿猴肾Vero细胞分离病毒,设计针对CVA16全VP1基因序列引物,采用RT-PCR扩增目的基因片段并测序;利用MEGA 7.0和Geneious 9.0.2等软件对全VP1序列进行分析.结果 共分离获得26株CVA16毒株,其中有8株KMB-17 分离株和 18 株 Vero 分离株.随机选取20株CVA16分离株进行分析,发现3株B1a和17株B1b基因亚型;其中17株CVA16 B1b分离株的核苷酸和氨基酸同源性分别为93.8％～100％和98.3％～100％,与国内其他分离株的核苷酸和氨基酸同源性为91.1％～99.2％和97.3％～99.0％;3株B1a分离株的核苷酸和氨基酸同源性为98.0％～98.1％和99.3％,与国内其他B1a分离株的核苷酸和氨基酸同源性为88.7％～98.7％和98.3％～99.7％;17株B1b分离株与其他3株B1a的核苷酸和氨基酸同源性为87.4％～88.4％和97.3％～98.7％.结论 2019年昆明地区流行的CVA16属于B1a和B1b基因亚型,以B1b为主,且均为中国大陆流行株.
Isolation and identification of Coxsackievirus A16 strains in Kunming,Yunnan Province in 2019 and analysis of genetic characteristics of VP1 gene
Objective To analyze the genetic characteristics of the entire VP1 gene of Coxsackievirus A16(CVA16)strains isolated from the feces of patients with hand,foot and mouth disease(HFMD)in Yunnan Province in 2019.Methods The virus was isolated from human embryonic lung diploid fibroblast(KMB-17)cells and African green monkey kidney(Vero)cells,and the primers for the complete VP1 gene sequence of CVA16 were designed.The target fragment was amplified by RT-PCR and sequenced;the complete VP1 sequence was analyzed by softwares such as MEGA 7.0 and Geneious 9.0.2.Results A total of 26 CVA16 strains were isolated,including eight KMB-17 isolates and 18 Vero isolates.Twenty CVA16 isolates were randomly selected for analysis,and three isolates were found to have B1a and 17 B1b genotypes;the nucleotide and amino acid homology of 17 CVA16 B1b isolates were 93.8％—100％and 98.3％—100％,and the nucle-otide and amino acid homology with other domestic isolates was 91.1％—99.2％and 97.3％—99.0％,respectively;the nucleotide and amino acid homology of the three B1a isolates was 98.0％—98.1％and 99.3％,and those with other domestic B1a isolates was 88.7％—98.7％and 98.3％—99.7％,respectively;17 B1b isolates and other three B1a isolates showed the nucleotide and amino acid homology of 87.4％—88.4％and 97.3％—98.7％.Conclusion The CVA16 prevalent in Kunming in 2019 belonged to Bla and B1b genotypes,with B1b as the main strain,and all of them were prevalent strains in the mainland of China.
NETL
NSTL
万方数据
