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: 10471460
• 负责人: Barry Ryan Lutz
• 金额: $ 75.62万
• 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 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10471460
• 关键词: Acute; Address; Adherence; Adopted; Aftercare; Antibodies; Area; Awareness; Biological Assay; Blood; CD4 Lymphocyte Count; Caring; Clinic; Clinic Visits; Complex; Country; Cytolysis; Decentralization; Detection; Developed Countries; Device Designs; Devices; Diagnosis; Diagnostic tests; Disclosure; Dropout; Drug resistance; Epitopes; Evaluation; Eye; Failure; Genomics; HIV; HIV Seronegativity; HIV Seropositivity; HIV diagnosis; Human; Human immunodeficiency virus test; Investigation; Lateral; Link; Methods; Monitor; Output; Paper; Patients; Performance; Persons; Phase; Plasmids; Policies; Population; Preparation; Protocols documentation; Public Health; RNA; RNA amplification; Reaction; Reagent; Reporting; Reproducibility; Research; Sampling; Schedule; Science; Services; Site; South Africa; Speed; Surface; System; Testing; Treatment Failure; United States; Viral; Viral Load result; Virion; Virus; Visual; Work; amplification detection; assay development; base; cellular targeting; clinical research site; cost; design; high risk; improved; innovation; internal control; isothermal amplification; novel; operation; point of care; point of care testing; pre-exposure prophylaxis; research clinical testing; sample archive; scale up; screening; self testing; sex; tool; viral detection; viral rebound

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 如果识别出艾滋病毒携带者，并通过治疗控制他们的病毒，艾滋病毒就可以得到有效的管理。 然而，许多艾滋病毒感染者不知道自己的艾滋病毒状况，因此需要增加检测的机会。在……里面 此外，由于耐药性或不遵守治疗计划，治疗可能会失败，而且测试是 需要确定治疗何时因这两个原因而失败。我们将开发一种检测方法来诊断艾滋病毒并检测 治疗失败，这是足够简单的，用户可以自己测试。 目的1.用于样品制备的艾滋病毒特异性捕获。我们将开发一种样品制备系统，直接 通过与病毒粒子表面结合的内源性人类抗体来靶向HIV病毒粒子，在 包膜和HIV病毒粒子表位。 目的2.核心分析方法的发展。我们将开发一种HIV的等温扩增分析方法，并设计一种 内部扩增对照。 目的3.快速、灵敏的HIV诊断方法。这项工作将建立在核心分析开发的基础上(目标2) 但增加了快速和灵敏的放大和检测的增强功能，并开发了芯纸设备。在……里面 此外，我们将检验一种假设，即检测敏感性可以通过艾滋病毒基因组靶标的贡献而增加。 从细胞碎片中分离出来。 目的4.病毒反弹的半定量检测。我们将以初步结果为基础，显示 基于阈值的检测，以开发半定量测试，报告单个阈值的是/否，以及 可以用眼睛来准备。 目的5.临床试验的设备设计、集成和放大(R33阶段)。为了达到这个目标，我们将完善 设计用于强健的操作、在内部评估性能和再现性，并向上扩展以生产设备 用于临床测试。 目的6.美国和南非的临床试验(R33阶段)。我们将对新鲜手指棒的测试进行评估 对美国西雅图和南非德班临床现场的血液进行评估，作为主要诊断 测试(1和2)，并与FDA批准的测试相比较，作为病毒学失败的测试(3和4)。