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[目的 ]建立一种 HIV- 1病毒基因亚型的分析方法 ,为了解福建省 HIV- 1各亚型的分布情况 ,提供方法学依据。 [方法 ]采集 HIV- 1感染者全血 ,分离单核细胞 (PBMCs) ,提取核酸后用套式 PCR扩增 HIV- 1env基因的核酸片段 ,与各标准亚型对照质粒的 PCR产物进行杂交 ,使同源部位配对形成异源双链二聚体 ,根据其在聚丙烯酰胺凝胶电泳中的泳动率确定其亚型。[结果 ]应用异源双链泳动分析法 (heteroduplex mobility assay,HMA) ,用于本省 4份 HIV-1样品的亚型分析 ,并均判定为 B亚型。[结论 ]异源双链泳动分析法是一种快速、价廉、特异和可靠的 HIV亚型分析方法 ,可用于 HIV- 1亚型分布的研究。
[Objective] To establish a method for the analysis of HIV-1 virus genotypes and to provide a methodological basis for understanding the distribution of HIV-1 subtypes in Fujian Province. [Methods] The whole blood of HIV-1 infected patients was collected and mononuclear cells (PBMCs) were isolated. Nucleic acid was extracted after nucleic acid amplification. The nucleic acid fragment of HIV-1env gene was amplified by nested PCR and hybridized with the PCR products of each standard subtype control plasmid. The homologous sites were paired to form heteroduplexes, and their subtypes were determined based on their mobility in polyacrylamide gel electrophoresis. [Results] The heteroduplex mobility assay (HMA) was used to analyze the subtypes of 4 HIV-1 samples in our province, all of which were classified as subtype B. [Conclusion] The heteroduplex mobility assay is a rapid, inexpensive, specific and reliable method for the analysis of HIV subtypes, which can be used to study the distribution of HIV-1 subtypes.