San Francisco VA Medical Center, San Francisco, CA 94121, USA.
J Virol Methods. 2011 Aug;175(2):261-5. doi: 10.1016/j.jviromet.2011.04.015. Epub 2011 Apr 22.
Although commercial tests are approved for detection of HIV-1 plasma viral loads ≥ 20 copies per milliliter (ml), only one specialized research assay has been reported to detect plasma viral loads as low as 1 copy/ml. This manuscript describes a method of concentrating HIV-1 virions from up to 30 ml of plasma, which can be combined with a commercial viral load test to create a widely available, reproducible assay for quantifying plasma HIV RNA levels less than 1 copy/ml. Using this pre-analytically modified assay, samples with a known level of 0.5 copy/ml were detected in 8 of 12 replicates (mean 0.47 copy/ml; 95% confidence interval (CI) 0.14-0.81 copy/ml) and samples with a known level of 1.0 copy/ml were detected in 13 of 13 replicates (mean 1.96 copy/ml; 95% CI 1.42-2.50 copy/ml). By concentrating virus from 30 ml of plasma, HIV RNA could be measured in 16 of 19 samples (84%) from 12 of 12 subjects (mean 2.77 copy/ml; 95% CI 0.86-4.68 copy/ml). The measured viral load correlated inversely (r = -0.78; p = 0.028) with the total duration of viral suppression (viral load<40 copies/ml).
虽然商业检测已被批准用于检测血浆中 HIV-1 病毒载量≥20 拷贝/毫升（ml），但仅有一项专门的研究检测方法可检测低至 1 拷贝/ml 的血浆病毒载量。本文描述了一种从多达 30 毫升血浆中浓缩 HIV-1 病毒粒子的方法，该方法可与商业病毒载量检测相结合，创建一种广泛可用且可重复的检测方法，用于定量检测低于 1 拷贝/ml 的血浆 HIV RNA 水平。使用这种预分析修改的检测方法，在 12 个重复实验中的 8 个（平均 0.47 拷贝/ml；95%置信区间[CI] 0.14-0.81 拷贝/ml）中检测到已知水平为 0.5 拷贝/ml 的样本，在 13 个重复实验中的 13 个（平均 1.96 拷贝/ml；95%CI 1.42-2.50 拷贝/ml）中检测到已知水平为 1.0 拷贝/ml 的样本。通过浓缩来自 30 毫升血浆的病毒，可在 12 名患者中的 11 名（16/19 样本；84%）的 19 个样本（平均 2.77 拷贝/ml；95%CI 0.86-4.68 拷贝/ml）中测量 HIV RNA。测量的病毒载量与病毒抑制的总持续时间呈负相关（r = -0.78；p = 0.028）。
