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

• 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
• 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

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 艾滋病毒载量的自我检测将大大减少新的感染，并为艾滋病毒阳性提供方法。 个人在自己家中监测病毒载量。在家里测试，然后分析 汇总数据还将帮助卫生保健提供者、项目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。开发自我检测HIV病毒的挑战 负荷测试是可怕的;例如，目前的实验室测试使用毫升静脉血提取在一个 临床设置。对于自我检测，仅使用微升手指针刺血液，其具有低HIV病毒拷贝 号码为了应对这些挑战，一个具有良好记录的跨学科科学家团队已经 该团队包括来自化学、生物医学工程和病毒学的专业知识。该团队已经 成功证明了使用流感病毒检测和抗病毒敏感性测量的能力， 在不到15分钟的时间里改造了血糖仪在这里，该策略的重点是检测艾滋病毒蛋白酶活性 使用重新设计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。在具体目标1（R61期）中，携带胱天蛋白酶-3酶原的 将产生HIV蛋白酶肽底物。暴露于HIV蛋白酶会导致活性半胱天冬酶-3 酶，其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。在具体目标2（R61相）中，介孔二氧化硅珠被封端， 用抗HIV蛋白酶抗体或肽底物将产生。无机催化剂将被嵌入 在珠子的孔里。暴露于HIV蛋白酶将使抗体或肽底物脱帽， 释放催化剂，其将与基底接触以产生电化学活性分析物， 用血糖仪检测。前两个具体目标的设计是相互独立的去风险 这个高风险高影响的项目具体目标1和2的成功执行将导致具体目标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。的结果 R33阶段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，使用再利用葡萄糖的新方法的原理证明 在急性期和病毒载量因抗病毒药物耐药性而增加时检测HIV的仪器， 将停止治疗。