Cham F, Heyndrickx L, Janssens W, Van der Auwera G, Vereecken K, De Houwer K, Coppens S, Whittle H, van der Groen G
Department of Microbiology, Institute of Tropical Medicine, Antwerp, Belgium.
AIDS Res Hum Retroviruses. 2000 Nov 20;16(17):1915-9. doi: 10.1089/08892220050195874.
A multiplex DNA PCR assay was developed for the simultaneous first-round amplification of HIV-1 gag and env fragments for the heteroduplex mobility assay (HMA). This assay was compared with the conventional amplification assay, using DNA extracted from PBMC samples from 30 HIV-1-seropositive individuals from The Gambia, who were enrolled between 1992 and 1997. From 27 of 30 (90%) samples both gag and env HMA fragments were amplified simultaneously. In one sample only the gag HMA fragment could be amplified by multiplex DNA PCR, and in two samples amplification was negative for both gag and env HMA in multiplex as well as the mono-DNA PCR. Of the 28 Gambian isolates subtyped by gag/env HMA or by sequencing and phylogenetic analysis, the majority (19 of 28; 68%) were intersubtype recombinant. Fifteen of 28 (53%) samples were circulating recombinant form (CRF) CRF02.AG variants. Two isolates clustering with the previously documented Gambian isolate GM4 (previously described as an env GC recombinant) are classified as gag A/env J recombinants.
开发了一种多重DNA PCR检测方法，用于同时首轮扩增HIV-1 gag和env片段，以进行异源双链迁移率分析（HMA）。使用从1992年至1997年招募的30名来自冈比亚的HIV-1血清阳性个体的外周血单核细胞（PBMC）样本中提取的DNA，将该检测方法与传统扩增检测方法进行比较。在30个样本中的27个（90%）中，gag和env HMA片段同时被扩增。在一个样本中，仅多重DNA PCR能够扩增gag HMA片段，而在两个样本中，多重及单重DNA PCR对gag和env HMA的扩增均为阴性。在通过gag/env HMA或测序及系统发育分析进行亚型分型的28株冈比亚分离株中，大多数（28株中的19株；68%）为亚型间重组体。28个样本中的15个（53%）为循环重组形式（CRF）CRF02.AG变体。两株与先前记录的冈比亚分离株GM4（先前描述为env GC重组体）聚类的分离株被分类为gag A/env J重组体。
