Analysis of HIV-1 drug resistant mutations by line probe assay and direct sequencing in a cohort of therapy naive HIV-1 infected Italian patients.
Analysis of HIV-1 drug resistant mutations by line probe assay and direct sequencing in a cohort of therapy naive HIV-1 infected Italian patients.
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DOI：
10.1186/1471-2180-1-30
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发表时间：
2001
期刊：
影响因子：
4.2
通讯作者：
La Placa M
中科院分区：
文献类型：
--
作者：
Re MC;Monari P;Bon I;Gibellini D;Vitone F;Borderi M;La Placa M
对新近感染HIV-1的个体进行的耐药性常规测定表明，耐药性变异体的传播可能增加。通过线性探针检测（利帕）和TruGene HIV-1检测试剂盒分析了27例未经治疗的HIV-1感染意大利患者的血浆样本，以检测赋予HIV-1耐药性的突变。这两项试验都揭示了与不同数量患者的耐药性相关的氨基酸取代。特别是，两个突变（K70 R和V118 I），可检测的利帕和测序分析，分别显示耐药NRTI在两个血浆样本。通过病毒序列分析，发现了至少三种突变，这些突变赋予了对NNRTI的耐药性，但不能通过商业利帕检测到，只能揭示与核苷逆转录酶类似物相关的突变。此外，大多数样品显示与蛋白酶抑制剂抗性相关的突变。值得注意的是，一个关键的突变，如V82 A（发现为混合物），和一些“不确定”的结果（9个样品），由于缺乏信号的线对应于一个特定的探针，只揭示了利帕，而一个可变数量的二次突变是可检测的只有TruGene HIV-1检测。即使需要进一步研究来确定不同检测方法对耐药菌株传播评价的影响，利帕可能在广泛的人群分析中有用，在这种情况下，需要在短时间内获得大量结果，而测序分析能够检测赋予NRTI和NNRTI耐药性的突变，可能被认为是一种更完整的检测方法，尽管更昂贵，技术上更复杂。
The routine determination of drug resistance in newly HIV-1 infected individuals documents a potential increase in the transmission of drug-resistant variants. Plasma samples from twenty seven therapy naive HIV-1 infected Italian patients were analyzed by the line probe assay (LIPA) and the TruGene HIV-1 assay for the detection of mutations conferring resistance to HIV-1. Both tests disclosed amino-acid substitutions associated with resistance in a variable number of patients. In particular, two mutations (K70R and V118I), detectable by LIPA and by sequencing analysis respectively, revealed resistance to NRTIs in two plasma samples. At least three mutations conferring resistance to NNRTIs, not detectable by commercial LIPA, able to reveal mutations associated only with nucleoside reverse transcriptase analogues, were disclosed by viral sequence analysis. Moreover, most samples showed mutations correlated with resistance to protease inhibitors. Remarkably, a key mutation, like V82A (found as a mixture), and some "indeterminate" results (9 samples), due the absence of signal on the lines corresponding to a specific probe, was revealed only by LIPA, while a variable number of secondary mutations was detectable only by TruGene HIV-1 assay. Even if further studies are necessary to establish the impact of different tests on the evaluation of drug-resistant strains transmission, LIPA might be useful in a wide population analysis, where bulk results are needed in a short time, while sequencing analysis, able to detect mutations conferring resistance to both NRTIs and NNRTIs, might be considered a more complete assay, albeit more expensive and more technically complex.