Large-scale evaluation of HIV-1 DNA drug resistance testing as a robust tool for clinical decision-making: A nationwide study in China

作者：Caihong Wu, Limin Zhang, Zhong Chen, W.D. Ma, Yanhua Fu, Ke Yang, Mei Liu, Yanjun Li, Xiaohong Chen, Mingjie Hou, Min Liu, Aihua Deng, Qingxia Zhao, Lukun Zhang, Quan Wang, Jun Peng, Yongli Li, Keji Deng, Jingsong Bai, Hai Long, Yaokai Chen, Hui Wang, Yun He, Li Jin, Jiahui Guo, Bianchuan Cao, Yizhi Cui, Min Wang, Tuofu Zhu, Jun Yao, Tong Wang · 发表于：Journal of Pharmaceutical Analysis · 年份：2025 · DOI：10.1016/j.jpha.2025.101513 · 被引用次数：3 · 研究领域：HIV/AIDS drug development and treatment、HIV Research and Treatment、Molecular Biology Techniques and Applications

Human immunodeficiency virus type 1 (HIV-1) drug resistance remains a major challenge in HIV/AIDS management, particularly in individuals with low-level viremia (LLV) where RNA-based drug resistance testing (DRT) often fails. Although HIV-1 DNA DRT represents a promising alternative, its clinical utility has been constrained by insufficient evidence. This nationwide study in China enrolled 9,428 people living with HIV (PLWH), analyzing 10,903 samples spanning a wide viral load (VL) spectrum. To improve RNA detection, an optimized primer design combined with an extracellular particle (EP)-HIV co-isolation technique was developed. We then evaluated the reproducibility of drug resistance mutation (DRM) profiles between paired RNA and DNA DRTs using Sanger sequencing (SS), with single-molecule sequencing employed to establish a dominant sequence threshold. Our findings demonstrated that primer optimization and EP-co-isolation significantly enhanced RNA amplification success. DRMs were prevalent across all VL strata. The combined concordance and degeneracy rates (C/D rates) (where multiple DNA DRMs included all RNA-derived DRMs) between RNA and DNA DRTs ranged from 90.4% to 100% in different gene regions, with higher discordance rates observed in the nucleoside reverse transcriptase inhibitor (NRTI) and non-NRTI (NNRTI) regions. Based on Stanford penalty scores across 25 antiretroviral drugs, the degeneracy group showed a 98.3% ± 1.7% interpretation agreement. Even within the disc...