Tatt I D, Barlow K L, Clewley J P
Hepatitis and Retrovirus Laboratory, Central Public Health Laboratory, London, United Kingdom.
J Virol Methods. 2000 Jun;87(1-2):41-51. doi: 10.1016/s0166-0934(00)00146-4.
A heteroduplex mobility assay (HMA) using 753 and 446 base pair (bp) amplicons of the p17/p24 region of the gag gene of HIV-1 has been developed and validated with reference clones and clinical samples representative of subtypes A, B, C, D, E, G, and H. There was complete concordance between the gag HMA assigned subtype and the subtype known from gag or env sequence data or env HMA. The heteroduplexes from both amplicons can be clearly resolved on either MetaPhor XR agarose or MDE polyacrylamide gels. The MetaPhor XR gel system was the more convenient and is the preferred choice for routine HMA subtyping. This gag HMA provides a rapid, simple and inexpensive method for subtyping HIV-1 based on a genomic region other than the commonly used env gene target. The incorporation of gag HMA into subtype determination algorithms should allow the detection of gag/env recombinant strains of HIV-1.
已开发出一种异源双链体迁移率分析（HMA）方法，该方法使用HIV-1 gag基因p17/p24区域的753和446碱基对（bp）扩增子，并通过代表A、B、C、D、E、G和H亚型的参考克隆及临床样本进行了验证。gag HMA确定的亚型与从gag或env序列数据或env HMA已知的亚型完全一致。两种扩增子的异源双链体在MetaPhor XR琼脂糖凝胶或MDE聚丙烯酰胺凝胶上均可清晰分辨。MetaPhor XR凝胶系统更方便，是常规HMA亚型分析的首选。这种gag HMA提供了一种基于不同于常用env基因靶点的基因组区域对HIV-1进行亚型分析的快速、简单且廉价的方法。将gag HMA纳入亚型确定算法应能检测出HIV-1的gag/env重组毒株。
