An ultrasensitive planar array p24 Gag ELISA to detect HIV-1 in diverse biological matrixes.

Human immunodeficiency virus-1 (HIV-1) persistence in the presence of antiretroviral therapy (ART) has halted the development of curative strategies. Measuring HIV persistence is complex due to the low frequency of cells containing virus in vivo. Most of the commercially available assays to date measure nucleic acid. These assays have the advantage of being highly sensitive and allow for the analysis of sequence diversity, intactness of the HIV genome or evaluation of diverse RNA species. However, these assays are limited in evaluating translational competent viral reservoirs. In here, we developed an ultrasensitive p24 ELISA that uses the Simoa planar array technology that can detect HIV-1 virions and HIV-1 infected cells with limit of detection similar to nucleic acid assays. Furthermore, the assay is optimized to measure very low levels of p24 in different biological fluids without a major loss of sensitivity or reproducibility. Our results demonstrate that the ‘homebrew’ planar p24 ELISA immunoassay is a broadly applicable new tool to evaluate HIV persistence in diverse biological fluids and cells.

在抗逆转录病毒疗法（ART）存在的情况下，人类免疫缺陷病毒1型（HIV - 1）的持续存在阻碍了治愈策略的发展。由于体内含病毒细胞的频率较低，测量HIV的持续存在情况较为复杂。到目前为止，大多数市售的检测方法都是检测核酸。这些检测方法具有高度灵敏的优点，并能够分析序列多样性、HIV基因组的完整性或评估不同的RNA种类。然而，这些检测方法在评估具有翻译能力的病毒储存库方面存在局限性。在此，我们开发了一种超灵敏的p24酶联免疫吸附测定（ELISA），它使用了单分子阵列（Simoa）平面阵列技术，能够检测HIV - 1病毒粒子和HIV - 1感染的细胞，其检测限与核酸检测方法相似。此外，该检测方法经过优化，可测量不同生物体液中极低水平的p24，且不会大幅损失灵敏度或重现性。我们的结果表明，这种“自制”的平面p24 ELISA免疫测定是一种广泛适用的新工具，可用于评估不同生物体液和细胞中HIV的持续存在情况。