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目的 运用异源双链泳动分析法 (HMA )进行人类免疫缺陷病毒 1(humanimmunodeficiencyvirus 1,HIV 1)基因亚型分析 ,了解福建省HIV 1亚型的流行情况。方法 收集细胞培养中HIV 1感染的外周血单个核细胞 (PBMC) ,聚合酶链反应 (PCR)扩增HIV 1膜蛋白基因(env)区的核酸片段 ,与标准亚型的对照质粒的PCR产物进行杂交 ,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 ,并进一步对标本进行序列测定和系统树分析。结果15份标本中 ,80 .0 0 %为E亚型 ,6 .6 7%为B亚型 ,2份不能确定其亚型。比较异源双链泳动法和序列测定法的结果 ,显示两种分析方法对HIV 1病毒基因分型有较高的一致性 ,其亚型一致率为86 .6 7%。结论 福建省流行的HIV 1毒株以E亚型为主。亚型分析法具有快速、简便、经济和高特异性的特点 ,可用作对HIV流行毒株进行长期监测的重要手段。
Objective To analyze the genotypes of human immunodeficiency virus 1 (HIV 1) using heteroduplex motility assay (HMA) to understand the prevalence of HIV 1 subtypes in Fujian Province. Methods Peripheral blood mononuclear cells (PBMCs) from HIV-1 infected cells were collected. The nucleic acid fragments of the env region of HIV 1 were amplified by polymerase chain reaction (PCR), and the PCR product of the control plasmid Were hybridized to form a heteroduplex dimer, and their subtypes were determined according to their mobility in polyacrylamide gel electrophoresis, and further sequencing and phylogenetic analysis of the samples were carried out. Results Of the 15 specimens, 80.0% were E subtypes, 6.7% were B subtypes, and 2 were undetermined subtypes. Comparison of heteroduplex motif and sequencing results showed that the two methods were highly consistent with HIV 1 genotyping, with a subtyping agreement of 86.67%. Conclusion The prevalence of HIV-1 strains in Fujian Province is mainly E subtypes. Subtype analysis is fast, easy, economical and highly specific and can be used as an important tool for the long-term monitoring of HIV-infected strains.