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：
10225439
负责人：
Suri Saranathan Iyer
金额：
$ 43.82万
依托单位：
GEORGIA STATE UNIVERSITY
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 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10225439
关键词：
AIDS/HIV problem Acute Address Adopted Anti-Retroviral Agents Antibodies Antigens Antiretroviral resistance Antiviral Agents 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 Head Health Personnel Home Human immunodeficiency virus test Incubated Individual Infection Inflammatory Bowel Diseases Influenza Intervention Laboratories Measures Medical Methods Monitor Neuraminidase Patients Peptide Hydrolases Peptides Performance Phase Plasma Policy Maker Predisposition Preparation Privacy Process Proteins Quality of life Reporter Reporting Research Reverse Transcriptase Polymerase Chain Reaction 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 base 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 pathogen point of care point-of-care diagnostics programs rapid detection self testing seropositive user-friendly virology virus culture volunteer

AIDS/HIV problem Acute Address Adopted Anti-Retroviral Agents Antibodies Antigens Antiretroviral resistance Antiviral Agents 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 Head Health Personnel Home Human immunodeficiency virus test Incubated Individual Infection Inflammatory Bowel Diseases Influenza Intervention Laboratories Measures Medical Methods Monitor Neuraminidase Patients Peptide Hydrolases Peptides Performance Phase Plasma Policy Maker Predisposition Preparation Privacy Process Proteins Quality of life Reporter Reporting Research Reverse Transcriptase Polymerase Chain Reaction 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 base 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 pathogen point of care point-of-care diagnostics programs rapid detection self testing seropositive user-friendly virology virus culture volunteer

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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.

项目摘要艾滋病毒病毒负荷的自我测试将显着减少新的感染，并提供艾滋病毒阳性的方法个人在自己家中监视病毒负荷。在家测试，然后分析汇总数据还将帮助医疗保健提供者，计划经理和政策制定者确定新的临床试验和其他干预措施的影响。发展自我测试的艾滋病毒病毒涉及的挑战负载测试是强大的；例如，当前的实验室测试使用在A中提取的静脉血的毫升临床环境。为了进行自我测试，仅使用了手指棒血的微粒，其HIV病毒拷贝较低数字。为了应对这些挑战，拥有良好记录的科学家跨学科团队已经组装；该团队包括化学，生物医学工程和病毒学的专业知识。团队有通过在不到15分钟的时间内重新利用的葡萄糖仪。在这里，该策略着重于检测HIV蛋白酶活性使用重新利用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本中，需要信号扩增。在特定的目标1（R61期）中，caspase-3蛋白酶轴承将产生HIV蛋白酶肽底物。接触HIV蛋白酶将导致活跃的caspase-3 酶将转换电化学不活性底物以产生电化学活性分析物用于检测和使用葡萄糖仪的资格。在特定的目标2（R61期）中，介孔二氧化硅珠盖了将产生抗HIV蛋白酶抗体或肽底物。无机催化剂将嵌入在珠子的毛孔内。暴露于HIV蛋白酶将开开抗体或肽底物，并释放催化剂，该催化剂将与底物到达，以产生电化学活性分析物的用葡萄糖仪检测。前两个具体目标的设计目的是彼此独立以发光风险这个高风险，高影响力项目。成功执行特定目标1和2将导致特定目标3（R33 阶段），这将涉及从培养和HIV阳性患者中测试病毒。来自将比较R33相，并与基于实验室的方法进行比较，包括高度精确的PCR 方法。在项目期结束时，使用重新利用葡萄糖的新方法的原理证明在急性期检测HIV的仪表以及由于抗病毒抗病性或将建立中止治疗。