Detection of residual plasma viremia in antiretroviral therapy (ART)-suppressed HIV-infected individuals is critical for characterizing the latent reservoir and evaluating the impact of cure interventions. Ultracentrifugation-based single-copy assays are sensitive but labor intensive. Fully automated replicate testing using a standard clinical viral load assay was evaluated as a high-throughput alternative for the quantification of low-level viremia. Four plasma samples from blood donors with acute HIV-1 infection and one viral culture supernatant were serially diluted into 25-ml samples to nominal viral loads ranging from 39 to <0.5 copies (cp)/ml. Each dilution was tested with 45 replicates (reps) using 0.5 ml/rep with the Aptima HIV-1 Quant assay. The nominal and estimated viral loads based on the single-hit Poisson model were compared, and a hybrid Poisson digital model for calibrated viral load estimation was derived. Testing performed using 45 reps on longitudinal plasma samples from 50 ART-suppressed individuals in the Reservoir Assay Validation and Evaluation Network (RAVEN) study cohort (range of 1 to 19 years of continuous ART suppression) showed a median viral load of 0.54 cp/ml (interquartile range [IQR], 0.22 to 1.46 cp/ml) and a 14% (95% confidence interval [CI], 9% to 19%) decline in viral load for each additional year in duration suppressed. Within the RAVEN cohort, the expected false-negative rate for detection at lower rep numbers using 9 and 18 reps was 26% and 14%, respectively. Residual plasma viremia levels positively correlated with cell-associated HIV RNA and DNA. The performance characteristics of the replicate Aptima assay support its use for quantifying residual plasma viremia to study the latent HIV reservoir and cure interventions.

中文翻译：

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复制Aptima分析以定量ART上个体的残留血浆病毒血症。

在抗逆转录病毒疗法（ART）抑制的HIV感染者中检测残留血浆病毒血症对于表征潜在储库和评估治疗干预的影响至关重要。基于超速离心的单拷贝测定法灵敏但劳动强度大。使用标准的临床病毒载量测定法进行的全自动复制测试被评估为定量低水平病毒血症的高通量选择。将来自患有急性HIV-1感染的献血者的四份血浆样品和一份病毒培养物上清液连续稀释到25 ml样品中，使标称病毒载量从39到<0.5拷贝（cp）/ ml。使用Aptima HIV-1 Quant测定法，以0.5 ml / rep进行45次重复（重复）测试每种稀释液。比较了基于一次命中的泊松模型的标称和估算病毒载量，并推导了用于校正病毒载量估计值的混合Poisson数字模型。在油藏试验验证和评估网络（RAVEN）研究队列（连续1到19年的连续抑制范围内）中，对来自50个被ART抑制的个体的纵向血浆样本进行了45次重复检测，结果显示病毒载量中位数为0.54 cp / ml （四分位间距[IQR]，0.22至1.46 cp / ml），且持续时间每延长一年，病毒载量下降14％（95％置信区间[CI]，9％至19％）。在RAVEN队列中，使用9次和18次重复进行较低重复次数检测时预期的假阴性率分别为26％和14％。残留血浆病毒血症水平与细胞相关的HIV RNA和DNA正相关。