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.

摘要 如果艾滋病毒感染者被识别出来，并且他们的病毒通过治疗得到控制，艾滋病毒就可以得到有效管理。 然而，许多艾滋病毒感染者并不知道自己的艾滋病毒状况，因此需要增加获得检测的机会。在 此外，由于耐药性或缺乏对治疗时间表的依从性，治疗可能失败， 需要确定治疗何时因任何原因而失败。我们将开发一种测试来诊断艾滋病毒， 治疗失败，这是简单的，足以让用户测试自己。 目标1.用于样品制备的HIV特异性捕获。我们将开发一种样品制备系统， 通过与病毒体表面结合的内源性人抗体、 包膜和HIV病毒体表位。 目标2.核心试验开发。我们将开发一种用于HIV的等温扩增检测方法，并设计一种 内部放大控制。 目标3.快速和灵敏的检测HIV诊断。这项工作将建立在核心检测开发（目标2）的基础上 但增加了快速和灵敏的扩增和检测的增强功能，并开发了核心纸装置。在 此外，我们将检验一个假设，即检测灵敏度可以通过HIV基因组靶点的贡献来提高 从细胞碎片。 目标4。病毒反弹检测的半定量试验。我们将根据初步结果， 基于阈值的检测，以开发半定量测试，该测试报告单个阈值的是/否， 可以用眼睛看。 目标5。用于临床试验的器械设计、集成和放大（R33阶段）。在本目标中，我们将完善 设计稳健的操作，内部评估性能和再现性，并扩大规模生产设备 用于临床试验。 目标6。在美国和南非进行临床试验（R33阶段）。我们将评估新鲜手指针刺的测试结果 在西雅图（美国）和德班（南非）的临床研究中心采集血液，以评价作为主要诊断的性能 测试（1和2）和作为病毒学失败的测试（3和4），通过与FDA批准的测试进行比较。