Section of Experimental and Clinical Pathology, Department of Biomedical Sciences and Human Oncology, University of Bari Aldo Moro, 70124 Bari, Italy.
Clinic of Infectious Diseases, Department of Biomedical Sciences and Human Oncology, University of Bari Aldo Moro, 70124 Bari, Italy.
Int J Mol Sci. 2022 Mar 1;23(5):2727. doi: 10.3390/ijms23052727.
Drug-resistance monitoring is one of the hardest challenges in HIV management. Next-generation sequencing (NGS) technologies speed up the detection of drug resistance, allowing the adjustment of antiretroviral therapy and enhancing the quality of life of people living with HIV. Recently, the NGS Sentosa SQ HIV Genotyping Assay (Vela Diagnostics) received approval for in vitro diagnostics use. This work is the first Italian evaluation of the performance of the Vela Diagnostics NGS platform, assessed with 420 HIV-1 clinical samples. A comparison with Sanger sequencing performance is also reported, highlighting the advantages and disadvantages of the Sentosa NGS assay. The precision of the technology was studied with reference specimens, while intra- and inter-assay reproducibility were evaluated for selected clinical samples. Vela Diagnostics' NGS assay reached an 87% success rate through 30 runs of analysis in a real-world clinical context. The concordance with Sanger sequencing outcomes was equal to 97.2%. Several detected mismatches were due to NGS's superior sensitivity to low-frequency variants. A high accuracy was observed in testing reference samples. Repeatability and reproducibility assays highlighted the good performance of the NGS platform. Beyond a few technical issues that call for further optimization, the key improvement will be a better balance between costs and processing speed. Once these issues have been solved, the Sentosa SQ HIV Genotyping Assay will be the way forward for HIV resistance testing.
耐药性监测是 HIV 管理中最具挑战性的难题之一。下一代测序（NGS）技术加速了耐药性的检测，使抗逆转录病毒治疗得以调整，并提高了 HIV 感染者的生活质量。最近，NGS Sentosa SQ HIV 基因分型检测试剂盒（Vela Diagnostics）获得了体外诊断用途的批准。本研究是对 Vela Diagnostics NGS 平台在意大利的首次评估，共评估了 420 份 HIV-1 临床样本。同时还报告了与 Sanger 测序性能的比较，突出了 Sentosa NGS 检测的优缺点。通过参考样本研究了该技术的精密度，同时针对选定的临床样本评估了内和间试验重复性。在实际临床环境中，通过 30 次分析运行，Vela Diagnostics 的 NGS 检测获得了 87%的成功率。与 Sanger 测序结果的一致性为 97.2%。几个检测到的不匹配是由于 NGS 对低频变异的更高敏感性所致。在测试参考样本时观察到了很高的准确性。重复性和重现性试验突出了 NGS 平台的良好性能。除了需要进一步优化的一些技术问题外，关键的改进将是在成本和处理速度之间取得更好的平衡。一旦解决了这些问题，Sentosa SQ HIV 基因分型检测试剂盒将成为 HIV 耐药性检测的未来趋势。
