Paper-based HIV self test

纸质 HIV 自检

基本信息

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
项目类别：
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

项目摘要

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：当前的艾滋病毒自我测试依赖于对病毒抗体的检测，因此不是在感染的早期阶段很有用。这些测定也不适合患者可能有病毒反弹的抗逆转录病毒疗法，作为预先存在的抗HIV抗体掩模病毒负荷的变化。相反，基于病毒RNA检测的当前HIV诊断很敏感，但太复杂且昂贵，无法自我测试。解决这个问题急需的需求，我们将开发第一个无酶的样本到艾滋病毒病毒载荷自我测试，指尖的血液被动地处理，导致孤立的 HIV基因组RNA。 RNA扩增策略将依靠发夹级联反应（HCR），在其他情况下，在其他情况下，在其他情况下，具有验证的单拷贝灵敏度的DNA纳米技术方法液滴实现。整个测定法将以廉价，一次性，基于纸张的消耗量。在R61阶段，我们建议：（1）构建我们拥有的方法先前验证的，优化全血的被动加工与纸基病毒裂解和RNA分离；（2）基于HCR的HIV基因组扩增的完整发展 RNA；（3）将样品处理和RNA扩增整合到单个消耗中；（4）获得有关艾滋病毒感染风险的患者的测定设计和可用性的反馈，并发展试点制造计划。在R33阶段，我们将（1）使用合同制造商的数量适合试点测试，以及（2）对测定的完整测试由1000名患有艾滋病毒感染的患者组成的队列，从参观该患者的患者中招募罗切斯特大学医学中心的急诊系。