Assessing the Suitability of Next-Generation Viral Outgrowth Assays to Measure Human Immunodeficiency Virus 1 Latent Reservoir Size.
Assessing the Suitability of Next-Generation Viral Outgrowth Assays to Measure Human Immunodeficiency Virus 1 Latent Reservoir Size.
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DOI：
10.1093/infdis/jiaa089
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发表时间：
2021-10-13
期刊：
影响因子：
--
通讯作者：
Busch MP
中科院分区：
文献类型：
--
作者：
Stone M;Rosenbloom DIS;Bacchetti P;Deng X;Dimapasoc M;Keating S;Bakkour S;Richman DD;Mellors JW;Deeks SG;Lai J;Beg S;Siliciano JD;Pagliuzza A;Chomont N;Lackman-Smith C;Ptak RG;Busch MP
人类免疫缺陷病毒（HIV）治疗干预措施的评价需要可靠和有效的量化复制能力的潜在水库。“经典的”定量病毒生长测定（QVOA）已被视为参考标准，尽管资源和劳动密集型。我们比较了6种“下一代”病毒生长检测方法，使用聚合酶链反应或超灵敏p24来评估它们作为QVOA可扩展代理的适用性。使用来自5名抗逆转录病毒治疗抑制的HIV感染受试者和1名HIV未感染对照的单个白细胞来源样本，将下一代QVOA与经典QVOA进行比较;每个实验室检测3份冷冻样本和1份新鲜样本的盲态批次。马尔可夫链蒙特卡罗方法估计额外的泊松变化在等分试样，批次和实验室水平。模型还估计了测试冷冻与新鲜样本的效果。下一代QVOA的变异估计值与QVOA相似。具有超灵敏度读数的测定报告了比经典QVOA更高的感染单位/百万值。对于下一代检测试剂盒，批内检测具有2.5倍的泊松外变异（95%可信区间[CI]，2.1-3.5倍）。实验室间变异使泊松外变异增加至3.4倍（95% CI，2.6-5.4倍）。冷冻储存没有显著改变每百万感染单位值（-18%; 95%CI，-52%至39%）。这些数据为使用下一代QVOA作为更费力的QVOA的代理提供了谨慎的支持，同时提供了更大的灵敏度和动态范围。根除策略中潜在储库的测量将受益于高通量和可扩展的测定。我们证明了经典和下一代定量病毒生长测定（QVOA）的变异估计值相似，灵敏度增强，支持使用下一代QVOA作为经典QVOA的代表，用于测量潜伏性人类免疫缺陷病毒库。
Evaluations of human immunodeficiency virus (HIV) curative interventions require reliable and efficient quantification of replication-competent latent reservoirs. The “classic” quantitative viral outgrowth assay (QVOA) has been regarded as the reference standard, although prohibitively resource and labor intensive. We compared 6 “next-generation” viral outgrowth assays, using polymerase chain reaction or ultrasensitive p24 to assess their suitability as scalable proxies for QVOA. Next-generation QVOAs were compared with classic QVOA using single leukapheresis-derived samples from 5 antiretroviral therapy–suppressed HIV-infected participants and 1 HIV-uninfected control; each laboratory tested blinded batches of 3 frozen and 1 fresh sample. Markov chain Monte Carlo methods estimated extra-Poisson variation at aliquot, batch, and laboratory levels. Models also estimated the effect of testing frozen versus fresh samples. Next-generation QVOAs had similar estimates of variation to QVOA. Assays with ultrasensitive readout reported higher infectious units per million values than classic QVOA. Within-batch testing had 2.5-fold extra-Poisson variation (95% credible interval [CI], 2.1–3.5-fold) for next-generation assays. Between-laboratory variation increased extra-Poisson variation to 3.4-fold (95% CI, 2.6–5.4-fold). Frozen storage did not substantially alter infectious units per million values (−18%; 95% CI, −52% to 39%). The data offer cautious support for use of next-generation QVOAs as proxies for more laborious QVOA, while providing greater sensitivities and dynamic ranges. Measurement of latent reservoirs in eradication strategies would benefit from high throughput and scalable assays. We demonstrated similar estimates of variation for classic and next-generation quantitative viral outgrowth assays (QVOAs), with enhanced sensitivity supporting the use of next-generation QVOAs as a proxy for classic QVOAs in measuring the latent human immunodeficiency virus reservoir.
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影响因子：
6.4
作者：
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通讯作者：
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影响因子：
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