The quantitative viral outgrowth assay (qVOA) is the gold standard for measuring inducible, replication-competent HIV-1. Using MOLT4-R5 and SupT1-R5 cell lines instead of allogeneic blasts and HIV-1 RNA detection rather than p24 enzyme-immunoassay (EIA) has been proposed to improve the sensitivity of the qVOA. It is unclear, however, how these alternative approaches affect qVOA performance. We compared three qVOAs methods across 15 persons with HIV-1 on suppressive antiretroviral therapy and found that the MOLT4-R5 method yielded a significantly higher proportion of p24-positive wells (42%) than both the allogeneic blast (29%) and SupT1-R5 (32%) assays. Additionally, 5 of 7 qVOAs that were negative by p24 EIA showed viral outgrowth by HIV-1 RNA quantification (>10-fold increase within 7 days). These findings reveal the potential for underestimation of the latent, inducible reservoir by qVOA depending on the target cells used and the measure of viral outgrowth. Use of MOLT4-R5 cells with both p24 EIA and HIV-1 RNA to detect viral outgrowth was the most sensitive method.

定量病毒生长试验 (qVOA) 是测量可诱导的、具有复制能力的 HIV-1 的金标准。已经提出使用 MOLT4-R5 和 SupT1-R5 细胞系而不是同种异体原始细胞和 HIV-1 RNA 检测而不是 p24 酶免疫测定 (EIA) 来提高 qVOA 的灵敏度。然而，尚不清楚这些替代方法如何影响 qVOA 性能。我们比较了 15 名 HIV-1 患者在抑制性抗逆转录病毒治疗中的三种 qVOA 方法，发现 MOLT4-R5 方法产生的 p24 阳性孔比例（42%）显着高于同种异体爆炸（29%）和 SupT1- R5 (32%) 检测。此外，p24 EIA 为阴性的 7 个 qVOA 中有 5 个通过 HIV-1 RNA 定量显示病毒生长（7 天内增加 > 10 倍）。这些发现揭示了 qVOA 可能低估潜在的、可诱导的储库，这取决于所使用的靶细胞和病毒生长的测量。使用具有 p24 EIA 和 HIV-1 RNA 的 MOLT4-R5 细胞来检测病毒生长是最敏感的方法。