V-OLA: point-of-care HIV viral load monitoring and drug resistance testing

V-OLA：即时 HIV 病毒载量监测和耐药性测试

基本信息

批准号：
10179314
负责人：
Barry Ryan Lutz
金额：
$ 70.61万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-14 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10179314
关键词：
Address Adherence Adoption Binding Biological Assay Blinded CLIA certified Chemistry Clinical Codon Nucleotides Consensus Counseling DNA Detection Devices Drug Targeting Drug resistance Engineering Equipment Failure Goals HIV HIV drug resistance HIV-1 Hour Individual Integrase Laboratories Lateral Ligation Link Medical Monitor Mutation Oligonucleotides Patients Peptide Hydrolases Performance Pharmaceutical Preparations Plasma Population Preparation Procedures Protocols documentation Quality Control RNA RNA-Directed DNA Polymerase Reagent Reflex action Regimen Reporting Research Resistance development Resources Risk Sampling Schedule Site Specimen System Temperature Testing Time Treatment Failure Tube Viral Load result Viral load measurement Virus Whole Blood Work acquired drug resistance antiretroviral therapy aquarium base clinically relevant cost design detection test detector digital drug testing experience implementation study innovation innovative technologies instrument internal control isothermal amplification laboratory equipment lateral flow assay mutant next generation sequencing operation point of care prevent rapid test resistance mutation scale up test strip transmission process usability

项目摘要

ABSTRACT We will develop the first test for detection of viral load (VL) and HIV drug resistance (DR) that is appropriate for low-resource settings (LRS). This new test is based on years of experience developing and testing the Oligonucleotide Ligation Assay (OLA) for HIV DR, as well as our recent work to develop OLA-Simple – a lab kit that reduces the time and complexity of OLA, but still requires laboratory equipment, laboratory one-way workflow to prevent contamination by amplicons, and hands-on operation of multiple steps. The proposed V- OLA test is a major advance over OLA-Simple that uses innovative technologies coordinated to give a low-cost, streamlined test suite appropriate for HIV patient management. We will develop rapid sample preparation from whole blood, semi-quantitative VL measurement at clinically-relevant thresholds (current WHO cut-off for treatment failure is >1000 copies/mL), and an integrated test cartridge to identify HIV DR mutations (goal to detect 10% mutant for patients with VL >1000 copies/mL). In Aim 1, we will develop chemistry and sample preparation format for HIV RNA isolation compatible with a rapid VL test. In Aim 2, we will develop the VL test using semi-quantitative isothermal amplification, including a simple low-cost heat block and detector. In Aim 3, we will develop core OLA chemistry including isothermal amplification of DR target regions, ligation on single- stranded amplicon, and dry reagents for amplification and OLA; we will test assays on different HIV subtypes. In Aim 4, we will develop the integrated DR detection device, including isothermal amplification and ligation with lateral flow strip detection. In Aim 5, we will combine the VF and DR tests into a coordinated system including refining hardware, scaling up fabrication of kit disposables, and develop a digitally-assisted system – called Aquarium – to provide step-by-step guided protocols, sample tracking, results interpretation, and scheduled quality control tests. We will test V-OLA on banked HIV plasma samples (HIV-1 subtypes of A, C, D, and AE). This level of testing is intended to provide preliminary evidence for in-depth studies on performance, usability, and implementation.

抽象的我们将开发第一个检测病毒载荷（VL）和HIV耐药性（DR）的测试低资源设置（LRS）。这项新测试基于多年的发展和测试经验 HIV DR的寡核苷酸连接测定法（OLA）以及我们最近开发Ola-Simple的工作 - 实验室套件这减少了OLA的时间和复杂性，但仍需要实验室设备，实验室单程工作流程以防止扩增子污染，并进行多个步骤操作。提议的V- OLA测试是与Ola-Simple的重大进步简化的测试套件适合艾滋病毒患者管理。我们将从全血，在临床上与临床相关的阈值处的半定量VL测量（当前截止治疗失败> 1000份/ml）和一个集成的测试弹药筒，以识别艾滋病毒DR突变（目标检测VL> 1000份/mL的患者的10％突变体。在AIM 1中，我们将开发化学和样品与快速VL检验兼容HIV RNA分离的制备格式。在AIM 2中，我们将开发VL测试使用半定量等温扩增，包括简单的低成本热块和检测器。在AIM 3中，我们将开发核心OLA化学，包括对DR目标区域的等温扩增滞留的扩增子和用于扩增和OLA的干试剂；我们将测试不同的HIV亚型的测定法。在 AIM 4，我们将开发集成的DR检测设备，包括等温扩增和连接横向流条检测。在AIM 5中，我们将将VF和DR测试结合到一个协调系统中精炼硬件，扩展套件的制造，并开发一个数字辅助系统 - 称为水族馆 - 提供逐步指导的协议，样本跟踪，结果解释和计划质量控制测试。我们将对储存的HIV血浆样品（A，C，D和AE的HIV-1亚型）测试V-OLA。这种测试水平旨在为性能，可用性，可用性研究提供初步证据和实施。