Rusconi S, La Seta Catamancio S, Sheridan F, Parker D
Istituto di Malattie Infettive e Tropicali, Universita' di Milano, Ospedale Luigi Sacco, Via GB Grassi 74, 20157, Milan, Italy.
J Clin Virol. 2000 Dec;19(3):135-42. doi: 10.1016/s1386-6532(00)00087-1.
The Murex-Innogenetics LiPA HIV-1 RT assay can be used to identify the presence of mutations of the reverse transcriptase gene at codons 41, 69, 70, 74, 184 and 215 of HIV-1, which have been shown to confer resistance to the nucleoside analogs Zidovudine (ZDV), Lamivudine (3TC), Didanosine (ddI) and Zalcitabine (ddC). The M184V mutation of the reverse transcriptase gene of HIV-1 has been associated with resistance to 3TC, ddC and ddI. This mutation has also been observed in patients receiving ZDV+ddC and ZDV+ddI. We used LiPA HIV-1 RT assay to identify the presence of either consensus methionine 184 or the mutant valine 184 with three groups of patients who were treated with ZDV/3TC, ZDV/ddI or ZDV/ddC combination therapy.
The aim of our study was to determine the viral genotype of patients who were considered to be failing therapy, by two ways: using sequencing and LiPA assays. In particular we were interested in establishing a possible correlation between these methods.
The study group consisted of a consecutive series of 33 patients with a treatment failure, 18 of whom received ZDV+3TC therapy, seven received ZDV+ddI and eight received ZDV+ddC therapy. We also examined a small cohort of seven seroconverters.
The M184V mutation was observed in 47.0% of patients receiving ZDV+3TC combination therapy but was not observed in either patient group receiving either ddI or ddC as co-therapy with ZDV. There was no evidence of the L74V mutation in our study group in either the ZDV/ddI or ZDV/ddC combination therapy group. We found the frequency of the K70R mutation to be higher in patients treated with ZDV/ddI (P=0.033) or ZDV/ddC (P=0.3) when compared with patients treated with ZDV/3TC.
The LiPA assay allowed for the rapid detection of wild type and amino acid variations at key positions conferring resistance to the most used antiviral RT inhibitors. This represented a rapid, quite sensitive, and simple genotyping test. For these reasons the LiPA assay proved to be useful in studying genetic resistance in large screenings, when key RT mutations could be useful in guiding an effective HIV-1 suppressing regimen.
Murex-Innogenetics LiPA HIV-1 RT检测法可用于鉴定HIV-1逆转录酶基因密码子41、69、70、74、184和215处突变的存在，这些突变已显示出对核苷类似物齐多夫定（ZDV）、拉米夫定（3TC）、去羟肌苷（ddI）和扎西他滨（ddC）具有耐药性。HIV-1逆转录酶基因的M184V突变与对3TC、ddC和ddI的耐药性有关。在接受ZDV+ddC和ZDV+ddI治疗的患者中也观察到了这种突变。我们使用LiPA HIV-1 RT检测法，对三组接受ZDV/3TC、ZDV/ddI或ZDV/ddC联合治疗的患者，鉴定是否存在共有的蛋氨酸184或突变的缬氨酸184。
我们研究的目的是通过两种方法确定被认为治疗失败患者的病毒基因型：使用测序和LiPA检测法。我们尤其感兴趣的是确定这些方法之间可能存在的相关性。
研究组由连续的33例治疗失败患者组成，其中18例接受ZDV+3TC治疗，7例接受ZDV+ddI治疗，8例接受ZDV+ddC治疗。我们还检测了一小群7例血清转化者。
在接受ZDV+3TC联合治疗的患者中，47.0%观察到M184V突变，但在接受ddI或ddC作为ZDV联合治疗的两组患者中均未观察到该突变。在我们的研究组中，ZDV/ddI或ZDV/ddC联合治疗组均未发现L74V突变的证据。我们发现，与接受ZDV/3TC治疗的患者相比，接受ZDV/ddI（P=0.033）或ZDV/ddC（P=0.3）治疗的患者中，K70R突变的频率更高。
LiPA检测法能够快速检测出对最常用抗病毒逆转录酶抑制剂产生耐药性的关键位置的野生型和氨基酸变异。这是一种快速、相当灵敏且简单的基因分型检测方法。由于这些原因，LiPA检测法在大规模筛查中研究基因耐药性时被证明是有用的，此时关键的逆转录酶突变可能有助于指导有效的HIV-1抑制方案。
