摘要
[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 study a genetic sub typing method of HIV 1 strains and understand the information of its sub types. [Methods] DNA fragments of HIV 1 env gene were amplified by nested PCR from uncultured mononuclear cells (PBMCs) obtained from HIV 1 infected individuals. The fragments were hybridized with the PCR product from reference plasmid of representative subtypes. Heteroduplex were then formed between the single stranded DNA from the two sources and were analyzed on the mobility of polyacrylamide gels. [Results] With heteroduplex mobility assay (HMA),four HIV 1 samples from Fujian were analyzed and determined as sub type B. [Conclusions] With advantages of high speed, low cost and high specificity, HMA is a reliable screening method for HIV 1 sub typing and may be used as the study on investigation of HIV 1 subtypes.
出处
《海峡预防医学杂志》
CAS
2000年第1期1-3,共3页
Strait Journal of Preventive Medicine
基金
福建省卫生厅青年科学研究基金
关键词
分子生物学
分子流行病学
HIV
HIV 1
genetic subtype
heteroduplex mobility assay (HMA)
molecular biology
molecular epidemiology
