HIV-specific target capture and quantitative isothermal amplification for acute HIV diagnosis and treatment monitoring

HIV 特异性目标捕获和定量等温扩增，用于急性 HIV 诊断和治疗监测

• 批准号: 10423662
• 负责人: Barry Ryan Lutz
• 金额: $ 74.6万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10423662
• 关键词: HIV; specific; target; capture; quantitative

Abstract HIV can be effectively managed if people with HIV are identified and their virus is controlled by therapy. However, many HIV-infected people do not know their HIV status, so increased access to testing is needed. In addition, therapy can fail due to drug resistance or lack of adherence to therapy schedules, and tests are needed to identify when therapy is failing for either reason. We will develop a test to diagnose HIV and detect treatment failure that is simple enough for users to test themselves. Aim 1. HIV-specific capture for sample preparation. We will develop a sample preparation system that directly targets HIV virions via endogenous human antibodies bound to the virion surface, human markers on the envelope, and HIV virion epitopes. Aim 2. Core assay development. We will develop an isothermal amplification assay for HIV and design an internal amplification control. Aim 3. Fast and sensitive detection for HIV diagnosis. This work will build on core assay development (Aim 2) but add enhancements for fast and sensitive amplification and detection and develop the core paper device. In addition, we will test a hypothesis that test sensitivity can be increased by contribution of HIV genomic targets from the cellular fraction. Aim 4. Semi-quantitative test for viral rebound detection. We will build upon preliminary results showing threshold-based detection to develop a semi-quantitative test that reports a yes/no for a single threshold and can be ready by eye. Aim 5. Device design, integration, and scale up for clinical testing (R33 Phase). In this Aim, we will refine the design for robust operation, evaluate performance and reproducibility in-house, and scale-up to produce devices for clinical testing. Aim 6. Clinical testing in the US and South Africa (R33 Phase). We will evaluate the tests on fresh fingerstick blood at clinical sites in Seattle, USA and Durban, South Africa to evaluate performance as a primary diagnostic test (1 and 2) and as a test for virologic failure (3 and 4) by comparison to FDA-cleared tests.

抽象的 如果鉴定出艾滋病毒患者，并且通过治疗控制了艾滋病毒，则可以有效地管理艾滋病毒。 但是，许多感染HIV的人不知道他们的艾滋病毒状况，因此需要增加对测试的访问。在 此外，由于耐药性或缺乏对治疗时间表的依从性，治疗可能会失败，并且测试是 需要确定何时出于任何原因进行治疗失败。我们将开发一项测试以诊断艾滋病毒并检测 治疗失败足以使用户对自己进行测试。 AIM 1。针对样品制备的HIV特异性捕获。我们将开发一个直接的样品制备系统 通过与病毒体表面结合的内源性人类抗体靶向HIV病毒体 信封和HIV病毒体表位。 目标2。核心测定开发。我们将开发用于艾滋病毒的等温扩增测定法和设计 内部扩增控制。 AIM 3。快速敏感的HIV诊断。这项工作将基于核心测定开发（AIM 2） 但是，为快速，敏感的放大和检测添加增强功能，并开发核心纸张设备。在 此外，我们还将检验一个假设，即通过HIV基因组靶标的贡献可以提高测试灵敏度 从细胞分数。 AIM 4。对病毒反弹检测的半定量测试。我们将基于展示的初步结果 基于阈值的检测以开发半定量测试，该测试报告单个阈值是/否，并且 可以眼睛准备好。 目标5。设备设计，集成和扩展进行临床测试（R33阶段）。在此目标中，我们将完善 设计可靠操作，评估内部性能和可重复性的设计，并扩大生产设备 用于临床测试。 AIM 6。在美国和南非（R33阶段）进行临床测试。我们将评估新鲜指法的测试 美国西雅图和南非德班的临床场所的血液评估表现为主要诊断 与FDA清除测试相比，测试（1和2），作为病毒衰竭（3和4）的测试。