Department of Microbiology, Immunology and Tropical Medicine, George Washington University, Washington, DC, USA.
Methods Mol Biol. 2024;2807:343-354. doi: 10.1007/978-1-0716-3862-0_23.
Assays to study HIV persistence are crucial to evaluate therapeutic strategies aimed toward an HIV cure. Several assays have been developed to date that rely on the measurement of nucleic acids. In recent years, the advancement of ultrasensitive technologies for the detection of proteins has improved our understanding of the role of translation-competent reservoirs in HIV persistence. In this chapter, we describe the development of an ultrasensitive p24 ELISA that uses planar array technology. This assay allows for the detection of HIV-1 p24 in the low fg/ml range in different biological matrixes, including cell lysates. This assay can be used to investigate the efficacy of latency reversing agents to reactivate HIV or to evaluate the persistence of translation-competent reservoirs in people living with HIV (PWH) in cells or diverse biological fluids.
用于研究 HIV 持续性的检测方法对于评估旨在实现 HIV 治愈的治疗策略至关重要。迄今为止，已经开发出了几种依赖于核酸测量的检测方法。近年来，用于检测蛋白质的超灵敏技术的进步提高了我们对翻译活性储库在 HIV 持续性中的作用的理解。在本章中，我们描述了一种使用平面阵列技术的超灵敏 p24 ELISA 的开发。该检测方法可以在不同的生物基质中（包括细胞裂解物）检测到低 fg/ml 范围内的 HIV-1 p24。该检测方法可用于研究潜伏逆转剂激活 HIV 的效果，或评估 HIV 感染者细胞或多种生物液中翻译活性储库的持续性。
