肠道病毒EV71型浙江分离株HZ08全基因组序列分析

目的了解浙江省流行的肠道病毒71型HZ08株的基因特征，研究肠道病毒71型的进化及变异状况。方法设计特异性引物，RT—PCR扩增HZ08株全基因，PCR产物纯化后克隆于T载体并进行序列测定和基因进化分析。结果HZ08株的全基因组由7405个核苷酸组成，编码2193个氨基酸。HZ08的5非编码区（UTR）、VPl、VP2、VP3、VP4、P2、P3、3非编码区（UTR）和全基因组的核苷酸与C4亚型的毒株序列较高，分别为96．2％～99．1％，92．6％～99．1％，92．7％～98．4％，88％～99．2％，93．8％～99．5％，89．4％～99．2％，91．4％～98．8％，87．8％～100％和91．1％～98．9％。HZ08株与其他亚型各区段的核酸同源性均低于90％。根据VPl基因和全基因序列进行种系统发生分析显示HZ08株与中国陆和台湾的病毒进化关系较近，与国际标准株以及东南亚毒株进化关系统较远。其中HZ08株与国内流行株BJ08和Fuyang08的进化关系最近。HZ08株与C4亚型相关氨基酸序列一致，VPl的抗原性表位发生了变异。结论HZ08病毒分离株为C4亚型，与Fuyang08和BJ08株进化途径相同，为肠道病毒7l型的基因组功能学和疫苗学研究奠定基础。

Complete genome sequence analysis of the enterovirus 71 HZ08 strain isolated in Zhejiang

To study the complete genome sequence of entrovirus 71 HZ08 strain gene isolated in Zhejiang province and explore its evolution. The total RNA was prepared from enterovirus 71 HZ08 strain and the RT-PCR products were cloned into T vector, sequenced and analyzed. The result of sequencing of the nucleotides showed that the complete gene of the HZ08 strain was 7405 bp in full length encoding 2193 amino acids. Data from sequences analysis showed that HZ08 shared 96.2~-99.1%, 92.6~ff0-99.1~,92.7~~-98.4~, 88~ff00-99.2~, 93.8~-99.5%0, 89.4~-99.2~~,91.4~- 98.8~,87.8~-100~ff00 and 91. 1~- 98.9~ in 5UTR, VP1,VP2, VP3,VP4, P2,P3,3UTR and complete genome with C4 subtype,respectively. HZ08 showed low nucleotides identity（~90~//0） with other subtypes. Phylogenetic trees established from alignment of the complete genome and VP1 region indicated that HZ08 shared the near genetic relationship with EVT1 isolated in china. HZ08 and other C4 subtype strains shared the same amino acids in 6 sites in VP1 region,which were associated with EV71 subtype. There was one mutation in VP1 antigen epitope. The HZ08 strain belonged to the same genogroup with Fuyang08 and Henan08 strains as C4 gene sub- types.