Van Laethem Kristel, Schrooten Yoeri, Covens Kris, Dekeersmaeker Nathalie, De Munter Paul, Van Wijngaerden Eric, Van Ranst Marc, Vandamme Anne-Mieke
Rega Institute for Medical Research, Katholieke Universiteit Leuven, Leuven, Belgium.
J Virol Methods. 2008 Nov;153(2):176-81. doi: 10.1016/j.jviromet.2008.07.008. Epub 2008 Sep 2.
Recently, the Food and Drug Administration (FDA) of the USA approved the first integrase inhibitor for inclusion in treatment regimens of HIV-1 patients failing their current regimens with multi-drug resistant strains. However, treatment failure has been observed during integrase inhibitor-containing therapy. Several mutational pathways have been described with signature mutations at integrase positions 66, 92, 148 and 155. Therefore, a genotypic assay for the amplification and sequencing of HIV-1 integrase was developed. The assay displayed a detection limit of 10 HIV-1 III(B) RNA copies/ml plasma. As the HIV-1 pandemic is characterised by a large genetic diversity, the new assay was evaluated on a panel of 74 genetically divergent samples belonging to the following genetic forms A, B, C, D, F, G, J, CRF01-AE, CRF02-AG, CRFF03-AB, CRF12-BF and CRF13-cpx. Their viral load ranged from 178 until >500,000 RNA copies/ml. The amplification and sequencing was successful for 70 samples (a success rate of 95%). The four failures were most probably due to low viral load or poor quality of RNA and not to subtype issues. Some of the sequences obtained from integrase inhibitor-naïve patients displayed polymorphisms at integrase positions associated with resistance: 74IV, 138D, 151I, 157Q and 163AE. The relevance of these polymorphisms in the absence of the signature mutations remains unclear.
最近，美国食品药品监督管理局（FDA）批准了首款整合酶抑制剂，用于纳入当前使用多药耐药菌株治疗方案失败的HIV-1患者的治疗方案中。然而，在含整合酶抑制剂的治疗过程中观察到了治疗失败的情况。已经描述了几种突变途径，整合酶第66、92、148和155位存在特征性突变。因此，开发了一种用于HIV-1整合酶扩增和测序的基因分型检测方法。该检测方法的检测限为每毫升血浆10个HIV-1 III(B) RNA拷贝。由于HIV-1大流行具有高度的遗传多样性，该新检测方法在一组74个基因差异样本上进行了评估，这些样本属于以下基因形式：A、B、C、D、F、G、J、CRF01-AE、CRF02-AG、CRFF03-AB、CRF12-BF和CRF13-cpx。它们的病毒载量范围从178到>500,000 RNA拷贝/毫升。70个样本的扩增和测序成功（成功率为95%）。这4次失败很可能是由于病毒载量低或RNA质量差，而不是亚型问题。一些从未接受过整合酶抑制剂治疗的患者获得的序列在与耐药相关的整合酶位置显示出多态性：74IV、138D、151I、157Q和163AE。在没有特征性突变的情况下，这些多态性的相关性尚不清楚。
