Towards point of care HIV detection using repurposed glucose meters

使用改造后的血糖仪进行艾滋病毒即时检测

基本信息

批准号：
10899285
负责人：
Suri Saranathan Iyer
金额：
$ 78.85万
依托单位：
UNIVERSITY OF MASSACHUSETTS LOWELL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10899285
关键词：
Acute Address Adopted Anti-Retroviral Agents Antibodies Antigens Antiretroviral resistance Articulation Biological Assay Biomedical Engineering Blood Buffers CASP3 gene Caregivers Chemistry Clinical Clinical Sensitivity Clinical Trials Data Data Aggregation Detection Development Diagnostic tests Disease Engineering Enzyme Precursors Enzymes Equipment Exposure to Feasibility Studies Fingers Goals Grant HIV HIV Protease HIV Seronegativity HIV Seropositivity HIV-1 HIV/AIDS Head Health Personnel Home Human immunodeficiency virus test Incubated Individual Infection Inflammatory Bowel Diseases Influenza Intervention Laboratories Licensing Measures Medical Methods Monitor Neuraminidase Pathogen detection Patients Peptide Hydrolases Peptides Performance Phase Plasma Policy Maker Predisposition Preparation Privacy Process Proteins Qualifying Quality of life Reaction Reporter Reporting Research Reverse Transcriptase Polymerase Chain Reaction Risk Sampling Scientist Sensitivity and Specificity Signal Transduction Silicon Dioxide Technology Testing United States National Institutes of Health Venous Viral Viral Drug Resistance Viral Load result Viremia Virion Virus Whole Blood Work World Health Organization acute infection burden of illness catalyst clinically relevant cytokine design enzyme activity feasibility testing field study glucose monitor high risk improved influenzavirus medical schools milliliter nanomolar novel particle point of care point-of-care diagnostics programs rapid detection self testing seropositive user-friendly virology virus culture volunteer

项目摘要

Project Summary Self-testing of HIV viral load would significantly reduce new infections and provide methods for HIV positive individuals to monitor viral load in the privacy of their own homes. Testing at home followed by analysis of the aggregate data would also assist health care providers, program managers and policy makers to determine the impact of new clinical trials and other interventions. The challenges involved in developing self-testing HIV viral load tests are formidable; for example, the current laboratory tests use milliliters of venous blood extracted in a clinical setting. For self-testing, only microliters of finger stick blood are used, which has low HIV viral copy numbers. To meet these challenges, an interdisciplinary team of scientists with a proven track record has been assembled; the team includes expertise from Chemistry, Biomedical Engineering and Virology. The team has successfully demonstrated the ability to detect influenza virus and measure antiviral susceptibility using repurposed glucose meters in less than 15 minutes. Here, the strategy focuses on detecting HIV protease activity using repurposed glucometers and correlating protease activity to viral load. Since low copy numbers are present in a sample, signal amplification is necessary. In specific aim 1 (R61 phase), a caspase-3 proenzyme bearing HIV protease peptide substrate will be produced. Exposure to HIV protease will result in an active caspase-3 enzyme, which will convert an electrochemically inactive substrate to produce an electrochemically active analyte for detection and qualification with glucometers. In specific aim 2 (R61 phase), mesoporous silica beads capped with anti-HIV protease antibody or peptide substrate will be produced. Inorganic catalysts will be embedded inside the pores of the beads. Exposure to HIV protease will uncap the antibodies or peptide substrate and release the catalysts, which will reach with a substrate to produce an electrochemically active analyte for detection with glucometers. The first two specific aims are designed to be independent of each other to de-risk this high risk, high impact project. Successful execution of specific aims 1 and 2 will lead to specific aim 3 (R33 phase), which would involve testing of the virus from culture and from HIV positive patients. The results from the R33 phase will be compared and contrasted to laboratory based methods including highly accurate PCR methods. At the end of the project period, proof of principle for a novel method that uses repurposed glucose meters to detect HIV during the acute phase and when the viral load increases due to antiviral resistance or discontinuing treatment will be established.

项目概要 HIV病毒载量的自我检测将显着减少新感染并为HIV阳性提供方法个人在自己家里的隐私中监测病毒载量。在家进行测试，然后进行分析汇总数据还将帮助卫生保健提供者、项目管理者和政策制定者确定新的临床试验和其他干预措施的影响。开发自检 HIV 病毒所面临的挑战负载测试非常艰巨；例如，目前的实验室测试使用从一个容器中提取的毫升静脉血。临床环境。对于自我检测，仅使用微升的指尖采血，其 HIV 病毒拷贝数较低数字。为了应对这些挑战，一支由具有良好业绩记录的跨学科科学家团队组成组装；该团队包括化学、生物医学工程和病毒学方面的专业知识。团队有成功证明了检测流感病毒和测量抗病毒敏感性的能力在 15 分钟内重新调整血糖仪的用途。在这里，该策略的重点是检测 HIV 蛋白酶活性使用重新调整用途的血糖仪并将蛋白酶活性与病毒载量相关联。由于存在低拷贝数在样品中，信号放大是必要的。在特定目标 1（R61 相）中，带有 caspase-3 酶原将产生HIV蛋白酶肽底物。接触 HIV 蛋白酶会产生活性 caspase-3 酶，它将转化电化学非活性底物以产生电化学活性分析物用于血糖仪的检测和鉴定。在特定目标 2（R61 相）中，介孔二氧化硅珠被封端与抗HIV蛋白酶的抗体或肽底物结合就会产生。将嵌入无机催化剂珠子的孔隙内。暴露于 HIV 蛋白酶会解开抗体或肽底物的盖子，释放催化剂，催化剂将到达底物以产生电化学活性分析物用血糖仪检测。前两个具体目标旨在相互独立，以降低风险这个高风险、高影响的项目。成功执行具体目标 1 和 2 将导致具体目标 3（R33 阶段），这将涉及从培养物和艾滋病毒阳性患者中检测病毒。结果来自 R33 相将与基于实验室的方法（包括高精度 PCR）进行比较和对比方法。在项目期结束时，使用重新利用的葡萄糖的新方法的原理证明在急性期以及由于抗病毒耐药性或病毒载量增加时检测艾滋病毒的仪表将确定停止治疗。