Cham F, Heyndrickx L, Janssens W, Vereecken K, De Houwer K, Coppens S, Van der Auwera G, Whittle H, van der Groen G
Department of Microbiology, Institute of Tropical Medicine, B-2000 Antwerp, Belgium.
AIDS Res Hum Retroviruses. 2000 Oct 10;16(15):1503-5. doi: 10.1089/088922200750006029.
The emergence of intersubtype recombinant HIV-1 isolates has made it imperative to analyze different regions of HIV-1 genomes. For this purpose a one-tube multiplex RT-PCR, coamplifying first-round amplicons that allow amplification of gag and env heteroduplex mobility assay (HMA) fragments from different HIV-1 group M isolates, was developed, starting with plasma samples. The multiplex RT-PCR assay is sensitive: 115 of 136 (84.5%) samples were positive for both gag and env, positive amplification of the gag fragment was observed in 130 of 136 (95.6%) samples, while for the env fragment 119 of 136 (87.5%) tested positive. The multiplex RT-PCR in combination with gag and env HMA makes large-scale HIV-1 subtyping fast, simple, and more economical.
HIV-1不同亚型重组毒株的出现使得分析HIV-1基因组的不同区域变得势在必行。为此，开发了一种单管多重逆转录聚合酶链反应（RT-PCR），从血浆样本开始，共同扩增第一轮扩增子，从而能够扩增来自不同HIV-1 M组毒株的gag和env异源双链迁移率分析（HMA）片段。该多重RT-PCR检测方法灵敏度高：136份样本中有115份（84.5%）的gag和env均呈阳性，136份样本中有130份（95.6%）观察到gag片段阳性扩增，而env片段检测阳性的有136份中的119份（87.5%）。多重RT-PCR结合gag和env HMA使得大规模HIV-1基因分型快速、简单且更经济。
