Assessment of intra-sample variability in HIV-1 DNA drug resistance genotyping.
Assessment of intra-sample variability in HIV-1 DNA drug resistance genotyping.
复制标题
HIV-1 DNA 耐药基因分型中样本内变异性的评估。
DOI：
10.1093/jac/dkab149
复制
发表时间：
2021
期刊：
影响因子：
--
通讯作者：
E. K. Alidjinou
中科院分区：
文献类型：
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作者：
Laurine Millière;L. Bocket;C. Tinez;O. Robineau;Nathalie Veyer;Fanny Wojciechowski;V. Lambert;A. Meybeck;T. Huleux;F. Ajana;D. Hober;E. K. Alidjinou
背景和目标
HIV-1耐药性检测可以在前病毒DNA中进行。细胞中病毒变体的非均匀分布可能会影响该方法的性能。我们使用下一代测序（NGS）方法评估了同一血液样本中HIV-1 DNA基因分型结果的变异性。
方法
对于每例入选患者，将单次静脉穿刺的血样分成5份1 mL等份，在同一运行中进行独立检测。使用实时PCR定量血液样本中的HIV-1 DNA，并使用Sentosa平台结合Sentosa SQ HIV基因分型测定进行NGS。
结果
共检测了12份样本（12例患者）的60份等分试样。中位年龄为45.50岁，所有患者均接受抗逆转录病毒治疗。有时可以在来自相同样本的等分试样之间的HIV-1 DNA定量中观察到显著的变异性，变异系数范围为23%至89%。20%临界值的耐药相关突变（RAM）分析发现，5、3和3例患者的相同样本等分试样之间的RAM谱在逆转录酶、蛋白酶和整合酶基因方面存在一些不一致性。具有较低截止值（10%）的分析显示了额外的突变，但没有改善样本内一致性。
结论
HIV-1 DNA耐药性检测结果存在样本内变异性，重复检测有时可能带来更多信息，但其临床影响程度仍需进一步调查。
BACKGROUND AND OBJECTIVES
HIV-1 drug resistance testing can be performed in proviral DNA. The non-homogenous distribution of viral variants in cells can impact the performance of this method. We assessed the variability of HIV-1 DNA genotyping results in the same blood sample using a next-generation sequencing (NGS) method.
METHODS
For each included patient, a blood sample from a single venipuncture was split into five 1 mL aliquots, which were independently tested in the same run. HIV-1 DNA was quantified in blood samples using real-time PCR, and NGS was performed with the Sentosa platform combined with the Sentosa SQ HIV genotyping Assay.
RESULTS
A total of 60 aliquots from 12 samples (12 patients) were tested. The median age was 45.50 years old, and all patients were treated with antiretrovirals. A significant variability can sometimes be observed in HIV-1 DNA quantification between aliquots from the same sample, with a coefficient of variation ranging from 23% to 89%. The analysis of resistance-associated mutations (RAMs) with a 20% cut-off found some discordances in RAMs profile between aliquots from the same sample for 5, 3 and 3 patients in the reverse transcriptase, protease and integrase genes, respectively. The analysis with a lower cut-off (10%) showed additional mutations, but did not improve the intra-sample concordance.
CONCLUSIONS
There is an intra-sample variability in HIV-1 DNA resistance test results, and repetition may sometimes bring additional information, but the extent of its clinical impact still requires further investigation.