Eugenin Eliseo A, Berman Joan W
Public Health Research Institute (PHRI), Rutgers University, 225 Warren Street, Newark, NJ, 07103, USA.
Department of Microbiology and Molecular Genetics, Rutgers University, 225 Warren Street, Newark, NJ, 07103, USA.
Methods Mol Biol. 2016;1354:265-79. doi: 10.1007/978-1-4939-3046-3_18.
Persistence of latent virus represents a major barrier to eradicating HIV even in the current antiretroviral therapy era. A critical limitation to eliminating these viral reservoirs is the lack of reliable methods to detect, quantify, and characterize cells harboring low levels of virus. However, recent work of several laboratories indicates that PCR and viral amplification based technologies underestimate or overestimate the size of the reservoirs. Thus, new technologies and methodologies to detect, quantify, and characterize these viral reservoirs are necessary to monitor and eradicate HIV. Recent developments in imaging technologies have enabled the development or improvement of detection protocols and have facilitated the identification and quantification of several markers with exquisite resolution. In the context of HIV, we developed new protocols for the detection of low amounts of viral proteins. In this chapter, we describe several antibody-based technologies for signal amplification to improve and detect low amounts of HIV proteins in cells, tissues, and other biological samples. The improvement in these techniques is essential to detect viral reservoirs and to design strategies to eliminate them.
即使在当前抗逆转录病毒治疗时代，潜伏病毒的持续存在仍是根除艾滋病毒的主要障碍。消除这些病毒储存库的一个关键限制是缺乏可靠的方法来检测、量化和表征携带低水平病毒的细胞。然而，几个实验室最近的研究表明，基于PCR和病毒扩增的技术低估或高估了储存库的规模。因此，需要新技术和方法来检测、量化和表征这些病毒储存库，以监测和根除艾滋病毒。成像技术的最新进展推动了检测方案的开发或改进，并有助于以高分辨率识别和量化多种标志物。在艾滋病毒的背景下，我们开发了检测少量病毒蛋白的新方案。在本章中，我们描述了几种基于抗体的信号放大技术，以改进和检测细胞、组织及其他生物样品中的少量艾滋病毒蛋白。这些技术的改进对于检测病毒储存库和设计消除它们的策略至关重要。
