广州市登革1型病毒基因组测序及系统进化分析

Complete genome sequencing and phylogenetic analysis of a Dengue virus type 1 strain isolated in Guangzhou

目的:对引起2002年广州登革热的登革1型病毒分离株(71/02GZ)进行全基因组测序和系统进化分析。方法:采用C6/36细胞培养71/02GZ,分别采用RT-PCR,5’RACE和3’RACE扩增基因组中编码区序列、5’和3’末端非编码区序列,PCR产物克隆到载体并测序,拼接成全基因组序列,分别基于E基因序列、E/NS1连接区240 nt、基因组中10 016 nt进行系统进化分析。结果:71/02GZ株基因组全长10 735 nt,两端非编码区(NCR)长度分别为94 nt和462 nt。基于E基因序列、E/NS1连接区240 nt、基因组中10 016 nt的进化分析结果显示71/02GZ株和2002年广州分离株(GZ01/02,GZ/218/2002),1995年广东分离株(GD23/95,GD01/95)、1995年广州分离株(GZ01/95)组成一个基因型;而1999年广东分离株(GD05/99),1997年广东分离株(GD14/97)和1980年广州分离株(GZ/80)属于另一个基因型;另外,71/02GZ株同1998年印度尼西亚分离株(98901530 DF DV-1-ID)的同源性最高。结论:71/02GZ株可能是印度尼西亚输入的。

Objective：To perform complete genome sequencing and phylogenetic analysis of a DENV-1 isolated（71/02GZ） strain which caused dengue fever in Guangzhou during 2002.Methods： Progagate 71/02GZ in C6/36 cells,the different fragments in viral genome including 5’terminal and 3’ terminal were amplified using RT-PCR,5’RACE and 3’RACE;PCR product was cloned to vector and then sequenced;Based on E gene nucleotide sequence、E/NS1 junction region 240 nt,10 016 nt of the genome,the phylogenetic relationship of 71/02GZ isolates and other DENV-1 isolates were analyzed, respectively. Results： The complete genome of 71/02GZ was 10735 nt, the 5＇ terminal and 3＇terminal NCRs were 94 nt and 462 nt, respectively. Phylogenetic analysis, based on E gene sequence, E/NS1 junction region 240 nt, 10 016 nt of the genome, showed that GD05/99 strain, GD14/97 strain and GZ/80 strain belonged to one genotype; 71/02GZ strain, GZ01/02 strain, GZ/218/ 2002 strain, GD23/95 strain, GD01/95 strain and GZ01/95 were the members of another genotype. In addition,71/02GZ strain was closely related with 98901530 DF DV-I-ID strain isolated in Indonesia in 1998. Conclusion： Strain of 71/02GZ may be imported from Indonesia.