Development of an HIV reverse transcriptase aptamer-based detection assay

描述（由申请人提供）：自1984年发现艾滋病毒以来，艾滋病毒的检测和定量一直是科学家和临床医生长期面临的问题。目前在现场或实验室有几种可用于筛查艾滋病毒感染的检测方法。它们通常分为几类：检测HIV RNA的基于pcr的检测，检测p24衣壳蛋白的ELISA检测，以及检测HIV逆转录酶（RT）的检测。还有检测艾滋病毒抗体的快速检测筛选试验，通常用作现场分析的初始筛选试验，因为它们高度敏感，只需几分钟即可完成。RT测定虽然在大多数情况下具有成本效益，但往往具有低灵敏度，这也是p24测定的缺点。基于聚合酶链反应的测定具有高灵敏度，但通常昂贵且耗时，并且需要专门的设备。这些检测也会像其他PCR检测一样受到污染。艾滋病领域将受益于一种廉价、敏感、定量、快速和简单的检测方法。一种在实验室环境中进行的少量病毒检测，可检测出药物筛选或突变分析可能导致的甚至非常低水平的感染。在这个应用程序中，我们提出了这样一个实验，使用我们在实验室开发的基于适配体的技术。我们的实验室已经开发出核酸适体，可以非常紧密地与HIV RT结合，并且能够检测到50-100个病毒颗粒。我们已经使用这些适体开始开发一种基于适体的RT检测分析（ARTA），它是敏感的，定量的，廉价的。我们将这项技术设想为下一代基于HIV rt的检测分析，并作为开发其他基于适配体的病原体检测分析的模型。

DESCRIPTION (provided by applicant): Detection and quantitation of HIV has been a longstanding issue for scientists and clinicians since the discovery of the virus in 1984. Currentl there are several assays available for screening for HIV infection in the field or the laboratory. They typically fall into a few categories: PCR-based assays that detect HIV RNA, ELISA assays that detect p24 capsid protein, and assays that detect HIV reverse transcriptase (RT). There are also rapid detection screening assays that detect antibodies to HIV and are typically used as the initial screening tests in field assays because of they are highly sensitive and take just minutes to complete. The RT assays though cost effective in most cases, tend to have low sensitivity which is also a drawback of p24 assays. PCR-based assays have high sensitivity but are typically costly and time consuming and require specialized equipment. These assays are also subject to contamination like other PCR assays. The HIV field would benefit from an inexpensive assay that is sensitive, quantitative, fast and simple. An assay that can be performed in a laboratory setting with small quantities of virus and could detect even very low levels of infection that might result from drug screens or mutational analysis. In this application we propose such an assay using aptamer-based technology that we have developed in our lab. Our lab has developed nucleic acid aptamers that can bind to HIV RT with very tightly and are capable of detecting as few as 50-100 virus particles. We have used these aptamers to begin to develop an Aptamer-based RT detection Assay (ARTA) that is sensitive, quantitative, and inexpensive. We envision this technology as the next generation of HIV RT-based detection assays and as a model for development of other aptamer-based pathogen detection assays.