意大利下一代测序中不同 HIV-1 抗药性解读工具的比较

Viruses Pub Date : 2024-09-06 DOI:10.3390/v16091422
Daniele Armenia, Luca Carioti, Valeria Micheli, Isabella Bon, Tiziano Allice, Celestino Bonura, Bianca Bruzzone, Fiorenza Bracchitta, Francesco Cerutti, Giovanni Maurizio Giammanco, Federica Stefanelli, Maria Addolorata Bonifacio, Ada Bertoli, Marialinda Vatteroni, Gabriele Ibba, Federica Novazzi, Maria Rosaria Lipsi, Nunzia Cuomo, Ilaria Vicenti, Francesca Ceccherini-Silberstein, Barbara Rossetti, Antonia Bezenchek, Francesco Saladini, Maurizio Zazzi, Maria Mercedes Santoro
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摘要

背景:下一代测序(NGS)正逐渐取代桑格测序用于 HIV 基因型耐药性检测(GRT)。本研究评估了在真实环境中不同 NGS-GRT 解释工具之间的一致性。方法:11 家意大利实验室使用 AD4SEQ HIV-1 Solution v2 商用试剂盒从病毒 RNA 中生成常规 NGS-GRT 数据。NGS 结果由试剂盒提供的 SmartVir 系统和两个在线工具(HyDRA Web 和 Stanford HIVdb)解读。当 HIVdb 9.5.1 算法中列出的每个 PR/RT/IN 耐药性相关位置的覆盖率大于 100 个读数(100×)时,NGS-GRT 被认为是有效的。结果在 629 个 NGS-GRT 中,SmartVir、HyDRA Web 和 HIVdb 的有效率分别为 75.2%、74.2% 和 70.9%。考虑到至少两种解读工具,463 个(73.6%)NGS-GRT 在耐药性分析中具有有效覆盖范围。有效样本的比例受病毒血症的影响,10% 的有效样本在不同解读工具之间表现出相当的一致性。结论这项意大利 NGS 耐药性检测调查表明,病毒血症水平和 HIV 亚型会影响 NGS-GRT 的覆盖率。在目前的 NGS-GRT 常规方法中,不同解读工具似乎只能可靠地检测到频率大于 10% 的突变。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of Different HIV-1 Resistance Interpretation Tools for Next-Generation Sequencing in Italy
Background: Next-generation sequencing (NGS) is gradually replacing Sanger sequencing for HIV genotypic drug resistance testing (GRT). This work evaluated the concordance among different NGS-GRT interpretation tools in a real-life setting. Methods: Routine NGS-GRT data were generated from viral RNA at 11 Italian laboratories with the AD4SEQ HIV-1 Solution v2 commercial kit. NGS results were interpreted by the SmartVir system provided by the kit and by two online tools (HyDRA Web and Stanford HIVdb). NGS-GRT was considered valid when the coverage was >100 reads (100×) at each PR/RT/IN resistance-associated position listed in the HIVdb 9.5.1 algorithm. Results: Among 629 NGS-GRT, 75.2%, 74.2%, and 70.9% were valid according to SmartVir, HyDRA Web, and HIVdb. Considering at least two interpretation tools, 463 (73.6%) NGS-GRT had a valid coverage for resistance analyses. The proportion of valid samples was affected by viremia <10,000–1000 copies/mL and non-B subtypes. Mutations at an NGS frequency >10% showed fair concordance among different interpretation tools. Conclusion: This Italian survey on NGS resistance testing suggests that viremia levels and HIV subtype affect NGS-GRT coverage. Within the current routine method for NGS-GRT, only mutations with frequency >10% seem reliably detected across different interpretation tools.
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