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应用套式PCR(nestedPCR)对24份HIV感染者外周血单核细胞(PBMC)样品进行扩增,从样品中获得了HIV1膜蛋白(env)基因的核酸片段,然后与已知各标准亚型的对照质粒的相同片段PCR产物进行杂交,使样品DNA与质粒DNA同源部位配对形成异源双链体,根据其在聚丙烯酰胺凝胶电泳中的泳动率可确定其亚型。并将这种异源双链泳动分析法(heteroduplexmobilityasay,HMA)的结果与DNA序列测定结果比较。实验证明,HMA是一种快速、简便、特异和可靠的HIV亚型的分析方法
Nucleic acid fragments of HIV-1 envelope protein (env) gene were obtained from 24 samples of peripheral blood mononuclear cells (PBMCs) of HIV-infected patients by nestedPCR and then compared with known Hybridization of the same PCR fragment of the control plasmid of each standard subtype results in the pairing of the sample DNA with the homologous portion of the plasmid DNA to form a heteroduplex which can be determined according to its mobility in polyacrylamide gel electrophoresis type. The result of this heteroduplex motility assay (HMA) was compared with that of DNA sequencing. Experiments show that HMA is a rapid, simple, specific and reliable method for the analysis of HIV subtypes