Towards point of care HIV detection using repurposed glucose meters

项目摘要 艾滋病毒病毒载量的自我检测将显著减少新的感染，并为艾滋病毒阳性提供方法 个人在自己家里监测病毒载量。在家中进行测试，然后分析 汇总数据还将帮助医疗保健提供者、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。开发自我检测艾滋病毒病毒所涉及的挑战 负荷测试是令人敬畏的；例如，目前的实验室测试使用的是从 临床环境。对于自我检测，只使用了微升的手指棒血，它的HIV病毒副本很低 数字。为了迎接这些挑战，一支由多个学科组成的科学家团队已经证明了这一点。 该团队包括来自化学、生物医学工程和病毒学的专业知识。这支队伍有 成功展示了检测流感病毒和测量抗病毒敏感性的能力 改装后的血糖仪只需不到15分钟。在这里，该战略的重点是检测艾滋病毒蛋白酶的活性 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。因为存在低拷贝数 在样品中，信号放大是必要的。在特定靶点1(R61期)，含有caspase-3酶原 将产生HIV蛋白酶肽底物。暴露在HIV蛋白酶下将导致caspase-3活性 酶，它将电化学反应不活跃的底物转化为电化学反应活性的分析物 用于血糖仪的检测和鉴定。在特定目标2(R61相)中，介孔二氧化硅小球被封顶 用抗HIV蛋白水解酶会产生抗体或多肽底物。无机催化剂将被嵌入 在珠子的毛孔里。暴露在HIV蛋白酶下会打开抗体或多肽底物的盖子， 释放催化剂，该催化剂将与底物接触以产生电化学活性分析物 用血糖仪检测。前两个具体目标被设计为相互独立以降低风险 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身上检测病毒。结果来自于 R33阶段将与基于实验室的方法(包括高精度的PCR)进行比较和对比 方法：研究方法。在项目期结束时，使用重新调整用途的葡萄糖的新方法的原理证明 在急性期和当病毒载量因抗病毒耐药性或 停止治疗将被确立。

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.