Department of Microbiology, Immunology and Tropical Medicine, George Washington University, Washington, USA.
Department of Microbiology and Immunology, Georgetown University School of Medicine, Georgetown University, Washington, USA.
Sci Rep. 2021 Dec 8;11(1):23682. doi: 10.1038/s41598-021-03072-7.
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 cell 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.
人类免疫缺陷病毒 1（HIV-1）在抗逆转录病毒疗法（ART）存在的情况下持续存在，这阻碍了治愈策略的发展。由于体内含有病毒的细胞频率较低，因此测量 HIV 持续性非常复杂。迄今为止，大多数商业上可用的检测方法都是测量核酸。这些检测方法具有高度灵敏性的优点，并且允许分析序列多样性、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翻译能力强的病毒储库方面存在局限性。在这里，我们开发了一种超灵敏的 p24 ELISA，该 ELISA 使用 Simoa 平面阵列技术，可以检测 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，可在不同的生物液中测量极低水平的 p24，而不会显著降低灵敏度或重现性。我们的研究结果表明，自制的平面 p24 ELISA 免疫分析是一种广泛适用的新工具，可用于评估不同生物液和细胞中的 HIV 持续性。
