Home Fingerprick Blood-Based HIV Self-Test For Quantitative Monitoring Of Viral Rebound

项目摘要/摘要 抗逆转录病毒治疗(ART)的进步显著降低了艾滋病毒的死亡率和发病率 将病毒载量(VL)降低到无法检测的水平。最近修订的世卫组织指南强烈建议常规 VL测试，以监控ART依从性，并将故障降至最低。为此，艾滋病毒自我检测，在这个过程中 想要了解艾滋病毒状况的个人私下采集样本，进行检测并解释结果， 已经成为一种赋权的方法。虽然核酸检测(NAT)很容易在集中式网络中使用 实验室用于病毒载量量化，但由于样本的原因，其在自我检测中的有效性尚未得到证明 加工和分析的复杂性。该项目旨在开发一种针对超小型USB的定量测试 检测病毒反弹的设备非常简单，外行人可以在美国进行自我测试 各州。 该项目的R61阶段将开发可以定量评估存在的整个基于血液的测试 HIV-1RNA的浓度低至1000拷贝/毫升，这是世卫组织推荐的检测阈值 治疗失败。在目标1中，我们将开发一种用于简化和自动化等离子体的一次性微流控芯片 从全血中分离和提取病毒RNA。在目标2中，我们将优化HIV-1 RT-LAMP检测和 探索最小拷贝数敏感度。在目标3中，我们将集成定量USB分析仪硬件 并开发操作简单、健壮的软件。在目标4中，我们将验证BSL-2中的原型测试 实验室在全血中加入了HIV-1血浆样本。 项目的R33阶段将评估HIV中的测试性能、可用性和利益相关者的需求 宾夕法尼亚州立好时医疗中心的综合护理计划。在目标5中，拟议的 对艾滋病毒感染者的检测将以标准的实验室方法为基准。在目标6中，我们将评估 用户对HIV VL自我检测的体验、态度和感知。 通过微流控芯片和USB分析仪的创新，我们预计这项测试将能够量化 HIV-1VL低至1000份/毫升，直接取自L的100微克手指刺血。这个项目的潜在影响 非常高。如果这个概念验证项目成功，它有可能显著提高 正在接受治疗的艾滋病毒感染者的治疗结果。潜在的转型能力保证了 与此项目相关的挑战。

Project Summary/Abstract The advancement of antiretroviral treatment (ART) has significantly reduced the HIV mortality and morbidity by reducing the viral load (VL) to undetectable levels. Recently revised WHO guidelines strongly recommend routine VL testing to monitor ART adherence and minimize failure. To this end, HIV self-testing, a process in which individual who wants to know HIV status collects a specimen, performs a test and interprets the result in private, has become an empowering approach. While nucleic acid testing (NAT) is readily available in centralized laboratories for viral load quantification, its availability in self-testing has not been demonstrated due to sample processing and assay complexity. This project aims to develop a quantitative test on an ultra-compact USB device to detect viral rebound that is simple enough for laypersons to test themselves in the United States. The R61 phase of the project will develop the whole blood-based test that can quantitatively assess the presence of HIV-1 RNA at concentrations as low as 1000 copies/ml, threshold recommended by WHO for determining treatment failure. In aim 1, we will develop a disposable microfluidic chip for streamlined and automated plasma separation and viral RNA extraction from whole blood. In aim 2, we will optimize the HIV-1 RT-LAMP assay and explore the minimum copy number sensitivity. In aim 3, we will integrate the quantitative USB analyzer hardware and develop software for easy and robust operation. In aim 4, we will validate the prototyped test in the BSL-2 lab using HIV-1 plasma samples spiked into whole blood. The R33 phase of the project will assess the test performance, usability and stakeholder needs within the HIV Comprehensive Care Program at Penn State Hershey Medical Center. In aim 5, the performance of the proposed test on HIV-infected patients will be benchmarked with standard laboratory methods. In aim 6, we will evaluate the user’s experience, attitude and perception of HIV VL self-testing. Through innovations in microfluidic chips and the USB analyzer, we anticipate the test would be able to quantify HIV-1 VL as low as 1000 copies/ml directly from 100 µl of finger prick blood. The potential impact of this project is very high. If this proof-of-concept project is successful, it has the potential to significantly enhance the treatment outcomes for individuals of HIV under therapy. The potential transformative capacity warrants the challenges associated with this project.