用于同时扩增HIV-1 M组gag和env异源双链迁移率分析片段的单管多重逆转录聚合酶链反应分析方法的开发。

Development of a one-tube multiplex reverse transcriptase-polymerase chain reaction assay for the simultaneous amplification of HIV type 1 group M gag and env heteroduplex mobility assay fragments.

作者信息

Cham F, Heyndrickx L, Janssens W, Vereecken K, De Houwer K, Coppens S, Van der Auwera G, Whittle H, van der Groen G

机构信息

Department of Microbiology, Institute of Tropical Medicine, B-2000 Antwerp, Belgium.

出版信息

AIDS Res Hum Retroviruses. 2000 Oct 10;16(15):1503-5. doi: 10.1089/088922200750006029.

DOI:10.1089/088922200750006029

PMID:11054263

Abstract

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.

摘要

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。