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目的测定广东省3株登革热2型病毒的全序列,探讨其来源及基因型。方法运用RT PCR法扩增来自广东省的登革热2型GD09/93、GD05/98和GD19/2001病毒株的全序列,采用遗传距离法构建进化树。结果3株登革热2型病毒的全基因组长度均为10 723 nt,5′及3′端各有一非编码区,结构基因和非结构基因位于基因组97~10 269 nt之间,共编码3391个氨基酸,读码结构完全相同。GD05/98与GD09/93、GD05/98与GD19/2001、GD09/93与GD19/2001之间的碱基序列同源性分别为93.3%、92.4%和97.6%,氨基酸序列同源性分别为96.7%、96.5%和98.5%。结论3株登革热2型病毒均对乳鼠致病,与对乳鼠不致病的参考株(DEN2-04株)比较共有18个氨基酸位点的差异引起极性或电荷变化,PrM-134、NS2A-153、NS4B-102所带电荷的变化对抗原性影响较大。GD05/98株与泰国分离株同为Ⅱ基因型,GD09/93和GD19/2001株与印度尼西亚、澳大利亚、台湾株分在Ⅳ基因型;表明我国登革热2型病毒有不同的基因型,而不同时期也存在同一种基因型传播。
Objective To determine the complete sequence of three dengue 2 viruses in Guangdong Province and discuss their origin and genotypes. Methods The complete sequence of dengue 2 GD09 / 93, GD05 / 98 and GD19 / 2001 strains was amplified by RT-PCR and the phylogenetic tree was constructed. Results The genomes of three Dengue virus type 2 viruses were all 10 723 nt in length and had a non-coding region at the 5 ’and 3’ end. The structural and nonstructural genes were located between 97 and 10 269 nt of the genome and encoded a total of 3391 amino acids , Reading structure exactly the same. The nucleotide sequence homologies between GD05 / 98 and GD09 / 93, GD05 / 98 and GD19 / 2001, GD09 / 93 and GD19 / 2001 were 93.3%, 92.4% and 97.6%, respectively. The amino acid sequence homologies were 96.7%, 96.5% and 98.5%. Conclusions All three Dengue virus type 2 viruses are pathogenic to neonatal rats. Compared with the non-pathogenic reference strain (DEN2-04 strain), a total of 18 amino acid residues cause changes in polarity or charge. PrM-134, NS2A-153, NS4B-102 charge brought about changes in the impact of antigenicity. GD05 / 98 isolates were the same as the two isolates from Thailand, GD09 / 93 and GD19 / 2001 isolates were from Indonesia, Australia and Taiwan. The genotypes of NDV GD05 / 98 strains were different in different genotypes, There is also the same genotype transmission.