异源双链泳动分析法在人类免疫缺陷病毒1基因亚型分析中的应用

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（human immunodeficiency virus-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，80.00%为E亚型，6.67%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86.67%。结论 福建省流行的HIV-1毒株以E亚型为主，亚型分析5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。

Application of heteroduplex mobility assay in genetic subtyping on human immunodeficiency virus type 1

Objective Using heteroduplex mobility assay(HMA) to subtype human immunodeficiency virus type 1(HIV 1) for the purpose of understanding HIV 1 subtype epidemic in Fujian province. Methods DNA fragments of HIV 1 env gene were amplified from peripheral blood mononuclear cell(PBMC) cocultures of HIV 1 infected individuals by nested polymerase chain reation(PCR). Heteroduplexs were formed through hybridizing PCR products from the samples and reference plasmid.According to the mobility of heteroduplexs in polyacrylamide gel electrophoresis, HIV 1 subtype from that sample was characterized and further confirmed by nucleotide sequencing analysis. Results Thirteen of 15( 86.67 %) samples were successfully subtyped by HMA,except 2 failures. Subtype E and B took up 80%(12/15) and 6.67 %(1/15) respectively. Results indicated a high concordance between HMA and nucleotide sequencing analysis and concordance rate was 86.67 %(13/15). Conclusions Subtype E appeared to be the major epidemic strain of HIV 1 in Fujian. HMA showed the characteristics of fastness, easiness, economie and with high specificity, and can be used in the survillence for the epidemic strain of HIV 1.