Clinical and Analytical Relevance of NNRTIs Minority Mutations on Viral Failure in HIV-1 Infected Patients
Clinical and Analytical Relevance of NNRTIs Minority Mutations on Viral Failure in HIV-1 Infected Patients
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DOI：
10.1002/jmv.23853
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发表时间：
2014-03-01
影响因子：
12.7
通讯作者：
Halfon, Philippe
中科院分区：
文献类型：
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作者：
Mohamed, Sofiane;Ravet, Sophie;Halfon, Philippe
本研究的目的是使用新的焦磷酸测序（PSQ）测定法评估少数变体的分析和临床相关性，并检测频率低于当前20%临床设定限值的次要变体。开发了一种用于检测和定量突变的PSQ方法，用于分析低于常规测序极限的人类免疫缺陷病毒（HIV）逆转录酶（RT）基因的14个密码子。10例患者在接受拉米夫定、替诺福韦、依非韦伦、奈韦拉平或依曲韦林一线治疗方案后出现病毒学失败（VF），以及10例对照患者无VF，均纳入本回顾性研究。使用桑格测序和PSQ方法评估基线血浆和VF时的血浆。检测次要序列变异的分析灵敏度为5%。在基线时，使用桑格测序和PSQ检测，在10/10例患者对照中未检测到少数变异，而2例治疗失败的患者具有标准基因分型未检测到的基线非核苷逆转录酶抑制剂（NNRTI）突变。在VF发生时，标准基因分型检测到10名VF患者中有4名发生突变，而PSQ检测到10名VF患者中有5名发生突变。临床上，在10%检测水平的少数突变可以通过焦磷酸测序有效地评估，并用作病毒群体进化的合适预测因子。这些特征允许更好地解释数据分析，这可以帮助临床医生提供合适的艾滋病毒治疗。医学病毒学杂志86：394-403，2014. (c)2013 Wiley Periodicals，Inc.
The objective of this study was to assess the analytical and clinical relevance of minority variants using a new pyrosequencing (PSQ) assay and to detect minor variants with frequencies below the current 20% clinical setting limit. A PSQ approach for detecting and quantifying mutations was developed for the analysis of 14 codons of the human immunodeficiency virus (HIV) reverse transcriptase (RT) gene below the limit of conventional sequencing. Ten patients who experienced virological failure (VF) after a first-line regimen of lamivudine, tenofovir, and either efavirenz, nevirapine, or etravirine, as well as 10 controls patients without VF, were included in this retrospective study. Baseline plasma and plasma from the time of VF were assessed using Sanger sequencing and PSQ methods. The analytical sensitivity for the detection of minor sequence variants is 5%. At baseline, no minority variant was detected in 10/10 patient controls using both the Sanger sequencing and PSQ assays, whereas, two patients who failed therapy had baseline non-nucleoside reverse transcriptase inhibitor (NNRTI) mutations that were not detected by the standard genotyping. At the time of VF, standard genotyping detected mutations in four out of the 10 VF patients, whereas, PSQ detected mutations in five out of the 10 VF patients. Clinically, minority mutations at a 10% level of detection can be assessed efficiently by pyrosequencing and used as a suitable predictor of the evolution of viral populations. These traits allow for a better interpretation of data analysis, which can help clinicians in providing a suitable treatment for HIV. J. Med. Virol. 86:394-403, 2014. (c) 2013 Wiley Periodicals, Inc.