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应用异源双链泳动分析法快速确定(HIV-1)基因亚型
RAPID GENETIC SUBTYPING OF HIV-1 USING A HETERODUPLEX MOBILITY ASSAY (HMA)
【摘要】 应用异源双链泳动分析法(Heterouduplex mobility assay,HMA) ,对来自我国云南、四川、新疆、浙江、广东、福建、天津7 省市73 份HIV 感染者样品,进行了HIV- 1 膜蛋白(env) 基因片段亚型分析。通过HMA 能确定亚型的有70 份,占样品总数的95-89 % (70/73) 。其中A 亚型2-86 % (2/70) ,F亚型2 .86 % (2/70) ,泰国B 亚型18 .58 % (13/70) ,欧美B 亚型8 .56 % (6/70) ,C亚型38 .57 % (27/70) ,E亚型28-57 % (20/70) 。为了检验结果的准确性和探讨3 份样品不能用HMA分型的原因,进一步对样品进行序列测定和系统树分析,并将两者结果的相互比较,两者对HIV- 1 分型显示出高度的一致性,其亚型一致率达95-89 % (70/73) 。而3 份样品不能确定亚型的原因主要是,其env 基因区段与相应的参考亚型质粒毒株相比变异太大。这提示,为了提高HMA分型的敏感性,应不断地选择代表性好的毒株构建参考亚型质粒。
【Abstract】 We investigated the DNA fragment diversity of HIV-1 env gene by means of heteroduplex mobility assay (HMA) genotyping. Samples of 73 HIV-1 infected individuals from Yunnan, Sichuan, Xinjiang, Zhejiang, Guangdong, Fujian and Tianjing of our country were studied. The HMA had successfully genotyped 95.89% (70/73) of these samples as follows: 2.86% (2/70) subtype A, 2.86% (2/70) subtype F, 18.58% (13/70) subtype Thailand B, 8.56% (6/70) subtype USA B, 38.57% (27/70) subtype C and 28.57% (20/70) subtype E. In order to test the reliability of these results and analyse the reason of failure to genotyping three samples by HMA, these strains were sequenced and phylogenetically analysed, and the results showed a perfect concordance with the HMA, the concordance rate was 95.89% (70/73). Reason of being unable to genotype three samples by HMA is that they were highly divergent subtype as compared to related reference strains. It suggests that, to improve reliability of HMA, the representative reference subtype plasmid selection must be continuously revised to cover viral diversification.
【Key words】 Human immunodeficiency virus type 1; Subtype; Heleroduplex mobility assay (HMA); Sequencing; Phylogenetic tree;
- 【文献出处】 病毒学报 ,CHINESE JOURNAL OF VIROLOGY , 编辑部邮箱 ,1999年04期
- 【分类号】R373
- 【被引频次】19
- 【下载频次】128