Development of an HIV reverse transcriptase aptamer-based detection assay

基于 HIV 逆转录酶适体的检测方法的开发

基本信息

批准号：
8846946
负责人：
JEFFREY J DESTEFANO
金额：
$ 7.6万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-12-01 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8846946
关键词：
Affinity Antibodies Antibody Formation Antigens Antiviral Agents Back Base Sequence Binding Biochemical Biological Assay Biosensor Capsid Proteins Categories Coupled Data Detection Development Diagnostic Enzyme-Linked Immunosorbent Assay Equipment Evolution Fluorescent Antibody Technique Gel HIV HIV Antibodies HIV Infections Health Human Individual Infection Laboratories Laboratory Scientists Ligands Methods Microscope Modification Molecular Biology Monitor Nucleic Acids Nucleic acid sequencing Nucleotides Pathogen detection Poly A Preclinical Drug Evaluation Proteins RNA RNA-Directed DNA Polymerase Radioactivity Scientist Scintillation Counting Signal Transduction Source Specificity Technology Testing Therapeutic Time Virion Virus Virus Diseases Virus Replication Western Blotting Work aptamer base cost cost effective drug resistant virus falls follow-up gel electrophoresis model development nanoparticle next generation oligo (dT)public health relevance rapid detection receptor research study screening tool

项目摘要

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.

描述（由申请人提供）：自1984年发现该病毒以来，对艾滋病毒的检测和定量一直是科学家和临床医生的一个长期问题。当前，有几种用于筛查该田间或实验室的HIV感染的测定法。它们通常属于几类：基于PCR的测定，检测HIV RNA，ELISA测定检测P24衣壳蛋白的测定以及检测HIV逆转录酶（RT）的测定。还有快速检测筛选测定法检测到与HIV的抗体，通常用作现场测定中的初始筛选测试，因为它们非常敏感，只需几分钟才能完成。 RT分析在大多数情况下具有成本效益，但敏感性较低，这也是P24分析的缺点。基于PCR的测定具有较高的灵敏度，但通常昂贵且耗时，需要专用设备。这些测定也像其他PCR分析一样受到污染。艾滋病毒场将受益于敏感，定量，快速和简单的廉价测定法。可以在具有少量病毒的实验室环境中进行的测定，甚至可以检测到可能由药物筛查或突变分析引起的非常低的感染水平。在此应用程序中，我们建议使用我们在实验室中开发的基于适体的技术提出这样的测定。我们的实验室已经开发了核酸适体，可以非常紧密地与HIV RT结合，并且能够检测到少于50-100的病毒颗粒。我们已经使用这些适体开始开发基于适体的RT检测测定法（ARTA），该测定敏感，定量和便宜。我们将这项技术设想为基于HIV RT的下一代检测分析，并将其作为开发其他基于适体病原体检测试验的模型。