Pont-of-use Acute HIV Infection Diagnostic for Substance Using Populations

针对药物使用人群的使用点急性 HIV 感染诊断

基本信息

批准号：
10794830
负责人：
Jacqueline Linnes
金额：
$ 28.93万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10794830
关键词：
3&apos Untranslated Regions Acceleration Acute Acute Hepatitis Acute Hepatitis C Antibodies Attitude Biological Assay Blood Blood capillaries Blood specimen Buffers Cessation of life Chronic Hepatitis C Clinic Clinical Cold Chains Continuity of Patient Care Cytolysis Detection Development Devices Diagnosis Diagnostic Diagnostic tests Drops Drug user Dryness Elements Exposure to Feedback Genome Goals HIV HIV Infections HIV diagnosis HIV-1 HIV/HCV Healthcare Heating Hepatitis C Hepatitis C Antibodies Hepatitis C Transmission Hepatitis C virus Human Human immunodeficiency virus test Individual Infection Infrastructure Injections Intervention Kentucky Laboratories Liver Cirrhosis Liver diseases Marketing Microfluidics Needle-Exchange Programs Nucleic Acid Amplification Tests Paper Pharmaceutical Preparations Pharmacy facility Plasma Population Preparation Process Proteins Protocols documentation RNA Rapid diagnostics Reaction Reagent Resistance Reverse Transcriptase Polymerase Chain Reaction Running Rural Rural Health Sampling Site Technology Testing Time Untranslated RNA Viral Viral Load result Virus Visual Waxes Whole Blood acceptability and feasibility acute infection anti-hepatitis C antibody detection barrier to care clinically relevant community based participatory research design detection limit health care availability high risk improved lateral flow assay novel novel strategies parent grant particle prevent prototype service providers sociocultural determinant substance abuse treatment substance use systemic barrier technology platform transmission process usability viron

项目摘要

Parent Grant: 1DP2DA051910 Parent Grant Title: Point-of-use Acute HIV Infection Diagnostic for Substance Using Populations Supplement Title: Expansion of Point-of-use Acute HIV Infection platform for Hepatitis C Virus An estimated 2.3 million individuals with HIV infection are co-infected with hepatitis C (HCV). In the US, people who use drugs (PWUD) are at high risk for both infections. HIV accelerates liver cirrhosis caused by HCV and contributes to the 350,000 deaths that occur due to HCV-related liver disease each year. Unfortunately, existing widely used rapid diagnostic tests (RDTs) for HCV do not detect the virus itself but instead detect only the anti-HCV antibodies produced by the infected individual. This antibody-based detection presents two challenges. First, host antibodies develop long after initial exposure to HCV which can allow for infections to remain undetected and potentially spread to other individuals. Second, anti-HCV antibodies remain even after HCV clearance and can result in false positive results that do not differentiate a current infection from a past infection. In contrast, test targeting the virus itself, such as nucleic acid amplification tests (e.g. RT-PCR), have the potential to detect both acute and chronic HCV infections. However, these assays typically require cold-chain storage of reagents, significant sample preparation, and extensive laboratory infrastructure to run. While PWUD are willing to get tested for HIV and HCV at non-traditional sites such as rural pharmacies (Duong et al 2019), current laboratory-based RT-PCR tests are unable to reach these individuals who are already underserved by traditional healthcare. Thus, there is an urgent need for a radically new approach to diagnose HCV among PWUD in order to engage infected individuals in the care continuum at the earliest stages of infection and prevent progression and transmission of HCV. In this supplement, we propose to adapt our current platform technology for acute HIV or order to to detect acute HCV thresholds above the clinically relevant 3000 HCV IU/mL (Reipold et al 2017) at point-of-use settings. Our ongoing DP2 is based on our handheld sample-to-answer technology, an automated microfluidic rapid autonomous analytical device (microRAAD). We have demonstrated that microRAAD is capable of detecting HIV virons directly from a whole blood samples. We incorporated dried amplification reagents and wax valves in paper-based substrates with resistive heating elements and low-power circuitry. By combining feedback-controlled heating of the RT-LAMP assay and wax valves with paper’s capillary flow, our assembled device automatically isolates viral particles from human blood, lysis the virus, amplifies HIV-1 RNA, and transports products to a detection zone with familiar visual lateral flow assay readout. This entire process requires only 3 simple steps for the user: first, a drop of blood is added to the sample pad, next, the user dispenses a buffer solution, and after a brief waiting period, the user reads the visual yes/no result. We have demonstrated that the fully integrated microRAAD detects as few as 10^5 HIV-1 vp/reaction directly from whole blood in under 90 minutes. In the ongoing DP2, we are working with local populations of PWUD and HIV service providers to evaluate feasibility, acceptability, and usability of the technology and to further reduce this detection to below 10^3 HIV vp/mL by combining p24 protein-based targeting and RT-LAMP together as well as improving clinical relevance via integration of an internal amplification control. Our next steps to adapt MicroRAAD to acute HCV detection will include: 1) Develop and optimize a novel assay for HCV detection targeting the conserved regions of the genome. For HCV assay design we will compare the 5’non-coding region (5’-NCR) and the 3’untranslated region (3’UTR) which have both been used in clinical HCV viral load assays. While the 5’-NCR is used in western assays, it’s known that the variability in this region is somewhat divergent from the sequences circulating worldwide. Therefore, we will also consider the 3’UTR for RT-LAMP design. We will generate at least two primers to each region and will compare the time to amplification (TTA), limit of detection (spiking in 0-10^7 IU/mL of synthetic RNA purchased from ATCC), and resistance to whole blood and plasma spiked into each sample at concentrations ranging from 0-20%. 2) Expand our community-based participatory research with PWUD and their service providers to evaluate HCV-focused detection at the point-of-use in concert with HIV detection. This will include performing feasibility and acceptability studies to evaluate the operational, clinical, and market needs specific for HCV detection by and for PWUD at the POU, evaluate how socio-cultural factors and systemic barriers to care among PWUD impact their HCV testing and integration into the current HIV care continuum and assess usability of device prototypes to determine the best operational protocols among stakeholders. This platform for point-of-use HCV/HIV detection could transform both testing and treatment of HIV and HCV among PWUD by bringing acute infection detection to substance abuse treatment facilities, needle exchanges, and injection drug clinics and will elucidate the targeted interventions needed to remove barriers to care. References: Reipold IE, Easterbrook P, Trianni A, Panneer N, Krakower D, Ongarello S, Roberts T, Miller V, Denkinger C. Optimising diagnosis of viraemic hepatitis C infection: the development of a target product profile. BMC Infect Dis. 2017 Nov 1;17(Suppl 1):707. doi: 10.1186/s12879-017-2770-5. PMID: 29143620; PMCID: PMC5688443. Duong M, Delcher C, Freeman PR, Young AM, Cooper HLF. Attitudes toward pharmacy-based HCV/HIV testing among people who use drugs in rural Kentucky. J Rural Health. 2022 Jan;38(1):93-99. doi: 10.1111/jrh.12564. Epub 2021 Mar 5. PMID: 33666274; PMCID: PMC8418619.

家长补助金：1DP2DA051910 家长资助名称：药物使用人群的使用点急性艾滋病毒感染诊断补充标题：针对丙型肝炎病毒的使用点急性 HIV 感染平台的扩展在美国，估计有 230 万艾滋病毒感染者同时感染丙型肝炎 (HCV)。使用药物 (PWUD) 是两种感染的高风险人群，HIV 会加速 HCV 引起的肝硬化，并导致不幸的是，每年有 35 万人因 HCV 相关肝病死亡。 HCV 诊断测试 (RDT) 不能检测病毒本身，而是仅检测产生的抗 HCV 抗体这种基于抗体的检测提出了两个挑战：首先，宿主抗体在很长时间后才产生。初次接触丙肝病毒可能导致感染未被发现并可能传播给其他人。其次，即使 HCV 清除后，抗 HCV 抗体仍然存在，并可能导致假阳性结果，而这些结果并不相反，针对病毒本身进行测试，例如核酸。扩增检测（例如 RT-PCR）有可能检测急性和慢性 HCV 感染。测定通常需要试剂的冷链储存、大量的样品制备和广泛的实验室同时 PWUD 愿意在农村等非传统地点进行艾滋病毒和丙肝病毒检测。药房（Duong et al 2019），目前基于实验室的 RT-PCR 检测无法覆盖到这些人传统医疗保健服务已经不足，因此迫切需要一种全新的方法来诊断 HCV。以便让感染者在感染和预防的最早阶段参与连续护理 HCV 的进展和传播。在本补充文件中，我们建议调整我们当前的急性艾滋病毒平台技术或检测急性丙型肝炎病毒我们正在进行的 DP2 阈值高于临床相关的 3000 HCV IU/mL（Reipold 等人 2017）。基于我们的手持式样品到答案技术，这是一种自动化微流体快速自主分析设备 (microRAAD)。我们已经证明 microRAAD 能够直接从全血中检测 HIV 病毒。我们将干燥的扩增试剂和蜡阀合并到带有电阻加热的纸基基底中。通过结合 RT-LAMP 检测和蜡阀的反馈控制加热。利用纸张的毛细管流动，我们组装的设备自动从人体血液中分离病毒颗粒，裂解病毒，扩增 HIV-1 RNA，并将产物运输至具有熟悉的视觉侧流测定读数的检测区域。整个过程对用户来说只需要 3 个简单的步骤：首先，将一滴血添加到样品垫上，然后，将用户分配缓冲液，短暂等待后，用户会读取视觉是/否结果。证明完全集成的 microRAAD 可以直接从全血中检测到少至 10^5 HIV-1 vp/反应在 90 分钟内，我们正在与当地残疾人士和艾滋病毒服务提供者合作，评估该技术的可行性、可接受性和可用性，并进一步将检测量降低到 10^3 HIV 以下 vp/mL 通过将基于 p24 蛋白的靶向和 RT-LAMP 结合在一起，并通过以下方式提高临床相关性集成内部放大控制。我们使 MicroRAAD 适应急性 HCV 检测的下一步措施将包括： 1) 开发并优化一种针对 HCV 基因组保守区域的 HCV 检测新方法。在设计中，我们将比较 5' 非编码区 (5'-NCR) 和 3' 非翻译区 (3'UTR)，它们都已被用于临床 HCV 病毒载量测定虽然 5’-NCR 用于西方测定，但众所周知，这种方法存在变异性。该区域与世界范围内流通的序列有些不同，因此，我们还将考虑 3’UTR。 RT-LAMP 设计。我们将为每个区域生成至少两个引物，并比较扩增时间 (TTA)，检测限（从 ATCC 购买的合成 RNA，加标浓度为 0-10^7 IU/mL），以及对全血和每个样品中加入的血浆浓度范围为 0-20%。 2) 与 PWUD 及其服务提供商一起扩大我们基于社区的参与性研究，以评估以 HCV 为重点的与 HIV 检测相结合的使用点检测这将包括执行可行性和可接受性。研究评估 POU 的 PWUD 检测 HCV 的具体操作、临床和市场需求，评估残疾人士中的社会文化因素和系统性护理障碍如何影响他们的丙型肝炎病毒检测和整合融入当前的艾滋病毒护理连续体并评估设备原型的可用性以确定最佳操作方案利益相关者之间。这个使用点 HCV/HIV 检测平台可以改变 HIV 和 HCV 的检测和治疗 PWUD 将急性感染检测引入药物滥用治疗机构、针头交换和注射药物诊所并将阐明消除护理障碍所需的有针对性的干预措施。参考： Reipold IE、Easterbrook P、Trianni A、Panneer N、Krakower D、Ongarello S、Roberts T、Miller V、Denkinger C. 优化病毒血症丙型肝炎感染的诊断：BMC Infect Dis 目标产品简介的开发。 11 月 1 日；17（增补 1）：707。PMID：29143620；PMC5688443。 Duong M、Delcher C、Freeman PR、Young AM、Cooper HLF 对基于药房的 HCV/HIV 检测的态度。肯塔基州农村地区吸毒者。J Rural Health 2022 年 1 月；38(1):93-99 doi: 10.1111/jrh.12564。 PMID：33666274；PMCID：PMC8418619。