Paper-based HIV self test

纸质 HIV 自检

• 批准号: 10555209
• 负责人: Benjamin L Miller
• 金额: $ 45.44万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-25 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10555209
• 关键词: Acceleration; Accident and Emergency department; Address; Biological Assay; Blood Volume; Blood capillaries; Caring; Catalytic DNA; Cellular Phone; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Contracts; Coupled; Cytolysis; DNA; Data; Data Set; Detection; Development; Devices; Diagnostic; Disease; Emergency department visit; Ensure; Enzymes; Feedback; Fingers; Fluorescence; Genetic Materials; Guidelines; HIV; HIV Antibodies; HIV Infections; Human immunodeficiency virus test; Humidity; Infection; Interview; Laboratories; Manufacturer; Masks; Medical center; Methods; Molecular; Monitor; Nanotechnology; Nucleic Acids; Paper; Patient Recruitments; Patients; Performance; Phase; Plasma; Plasma Cells; Porosity; Preparation; Process; Production; RNA; RNA amplification; Reaction; Reader; Research; Risk; Sampling; Sensitivity and Specificity; Source; Specificity; Structure; Temperature; Testing; Universities; Validation; Viral; Viral Load result; Virion; Virus; Whole Blood; Work; amplification detection; antibody detection; antiretroviral therapy; cohort; design; detection assay; detection limit; genomic RNA; innovation; instrumentation; manufacture; manufacturing test; novel strategies; operation; pathogenic virus; performance tests; pilot test; prototype; self testing; stability testing; success; usability; viral RNA; viral detection; viral rebound; whole genome

Abstract: Current self-tests for HIV rely on detection of antibodies to the virus, and thus are not useful during the early stages of infection. These assays are also not appropriate for patients on antiretroviral therapy who may be at risk for viral rebound, as pre-existing anti-HIV antibodies mask changes in viral load. Conversely, current HIV diagnostics based on detection of viral RNA are sensitive, but too complex and expensive to be appropriate for self-testing. To address this critical need, we will develop the first enzyme-free sample-to-answer HIV viral load self-test, in which a fingerstick volume of blood is passively processed leading to amplification of the isolated HIV genomic RNA. The RNA amplification strategy will rely on hairpin cascade reactions (HCR), a DNA nanotechnology approach with proven single-copy sensitivity in other contexts, in a sessile droplet implementation. The entire assay will be implemented in an inexpensive, disposable, paper-based consumable. In the R61 phase, we propose to: (1) building on methods we have previously validated, optimize passive processing of whole blood coupled with paper-based viral lysis and RNA isolation; (2) complete development of HCR-based amplification of HIV genomic RNA; (3) integrate sample processing and RNA amplification into a single consumable; and (4) obtain feedback on assay design and usability from patients at risk for HIV infection, and develop a pilot manufacturing plan. In the R33 phase, we will (1) scale production of the assay with contract manufacturers in quantities suitable for pilot testing, and (2) complete testing of the assay with a cohort of 1000 patients at-risk for HIV infection, recruited from patients visiting the Emergency Department at the University of Rochester Medical Center.

翻译后摘要：目前的自我测试艾滋病毒依赖于检测抗体的病毒，因此不是 在感染的早期阶段是有用的。这些测定也不适用于 抗逆转录病毒治疗可能存在病毒反弹风险，因为预先存在抗HIV抗体 掩盖病毒载量的变化相反，目前基于病毒RNA检测的HIV诊断 是敏感的，但太复杂和昂贵，不适合自我测试。为了解决这个 关键的需要，我们将开发第一个无酶样品回答艾滋病毒载量自我测试，在 其中手指针刺体积的血液被被动处理，导致分离的 HIV基因组RNA。RNA扩增策略将依赖于发夹级联反应（HCR）， 一种DNA纳米技术方法，在其他情况下具有已证明的单拷贝灵敏度， 液滴实现。整个测定将在廉价的、一次性的、 纸质耗材。在R61阶段，我们建议：（1）建立在我们已有的方法基础上 先前已验证，优化全血的被动处理加上纸基病毒 裂解和RNA分离;（2）完成基于HCR的HIV基因组扩增 （3）将样品处理和RNA扩增集成到单个消耗品中;以及（4） 从有HIV感染风险的患者那里获得关于检测设计和可用性的反馈，并开发 一个试验性的生产计划在R33阶段，我们将（1）使用以下试剂盒进行规模生产 合同制造商，数量适合于中试测试，以及（2）完成分析测试 与1000名艾滋病毒感染风险患者的队列，从访问的患者招募， 罗切斯特大学医学中心急诊科。