Paper-based HIV self test

纸质 HIV 自检

• 批准号: 10373830
• 负责人: Benjamin L Miller
• 金额: $ 47.2万
• 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/10373830
• 关键词: Accident and Emergency department; Address; Biological Assay; Blood Volume; Blood capillaries; Caring; Catalytic DNA; Cellular Phone; Centers for Disease Control and Prevention (U.S.); 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; Fluorescence; Genetic Materials; Gold; Guidelines; HIV; HIV Antibodies; HIV Infections; Human immunodeficiency virus test; Humidity; Infection; Interview; Laboratories; Manufacturer Name; Masks; Medical center; Methods; Molecular; Monitor; Nanotechnology; Nucleic Acids; Paper; Patient Recruitments; Patients; Performance; Phase; Plasma; Plasma Cells; Preparation; Process; Production; RNA; RNA amplification; Reaction; Reader; Research; Risk; Sampling; Sensitivity and Specificity; Source; Specificity; Structure; Temperament; Temperature; Testing; Universities; Validation; Viral; Viral Load result; Virion; Virus; Whole Blood; Work; amplification detection; antibody detection; antiretroviral therapy; base; cohort; design; detection assay; detection limit; genomic RNA; innovation; instrumentation; 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.

摘要：目前的艾滋病毒自我检测依赖于病毒抗体的检测，因此不可行。 在感染的早期阶段很有用。这些测定也不适合患者 接受抗逆转录病毒治疗的人可能面临病毒反弹的风险，因为已有抗 HIV 抗体 掩盖病毒载量的变化。相反，当前的 HIV 诊断基于病毒 RNA 检测 很敏感，但过于复杂且昂贵，不适合自测试。为了解决这个问题 迫切需要，我们将开发第一个无酶样本来回答 HIV 病毒载量自检， 被动处理指尖血量，导致分离的扩增 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，在固着中具有经过验证的单拷贝敏感性 液滴实施。整个测定将在一个廉价的、一次性的、 纸质耗材。在 R61 阶段，我们建议：（1）基于我们现有的方法 先前经过验证，优化全血的被动处理与纸基病毒相结合 裂解和 RNA 分离； (2) 完成基于HCR的HIV基因组扩增 核糖核酸； (3) 将样品处理和RNA扩增集成到单一耗材中；和（4） 从有 HIV 感染风险的患者那里获取有关检测设计和可用性的反馈，并制定 试点制造计划。在 R33 阶段，我们将 (1) 扩大检测规模生产 合同制造商的数量适合进行中试测试，以及 (2) 完成测定测试 一项由 1000 名有感染 HIV 风险的患者组成的队列，是从就诊的患者中招募的 罗切斯特大学医学中心急诊科。