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

• 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
• 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' 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

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.

家长资助：1DP 2DA 051910 家长资助项目：用于物质使用人群的使用点急性HIV感染诊断 补充标题：扩展丙型肝炎病毒的使用点急性HIV感染平台 估计有230万艾滋病毒感染者同时感染丙型肝炎（HCV）。在美国， 使用药物（PWUD）是这两种感染的高风险。HIV加速HCV引起的肝硬化， 每年有350，000人死于与HCV相关的肝脏疾病。目前广泛使用的快速 HCV诊断试验（RDTs）不能检测病毒本身，而只能检测产生的抗HCV抗体 被感染的个体。这种基于抗体的检测提出了两个挑战。首先，宿主抗体在 最初暴露于HCV可使感染保持未被发现并可能传播给其他个体。 第二，即使在HCV清除后，抗HCV抗体仍然存在，并可能导致假阳性结果， 区分当前感染和过去感染。相反，针对病毒本身的测试，如核酸 扩增测试（例如RT-PCR）具有检测急性和慢性HCV感染的潜力。但这些 测定通常需要试剂的冷链储存、大量的样品制备和大量的实验室操作。 基础设施运行。虽然PWUD愿意在农村等非传统地点接受艾滋病毒和丙型肝炎病毒检测， 药房（Duong et al 2019），目前基于实验室的RT-PCR检测无法达到这些个体， 传统的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 为了使受感染者在感染的最早阶段接受连续护理， HCV的传播和传播。 在本补充中，我们建议调整我们目前的急性艾滋病毒平台技术，以检测急性HCV 在使用点设置下，阈值高于临床相关的3000 HCV IU/mL（Reipold et al 2017）。我们正在进行的DP 2 是基于我们的手持样品回答技术，一种自动化的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 样品我们将干燥的扩增试剂和蜡阀在纸基基板与电阻加热 元件和低功耗电路。通过将RT-LAMP测定和蜡阀的反馈控制加热相结合， 利用纸的毛细流动，我们组装的装置自动从人体血液中分离病毒颗粒，溶解病毒， 扩增HIV-1 RNA，并将产物运输到具有熟悉的可见侧向流测定读数的检测区。 对于用户来说，整个过程只需要3个简单的步骤：首先，将一滴血液添加到样品垫，然后， 用户分配缓冲溶液，短暂等待后，用户读取视觉是/否结果。我们有 研究表明，完全集成的microRAAD直接从全血中检测到10^5 HIV-1 vp/反应， 不到90分钟在正在进行的DP 2中，我们正在与当地的PWUD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，并进一步将检测结果降至10^3以下 通过将基于p24蛋白的靶向和RT-LAMP结合在一起，以及通过 内部放大控制的集成。 我们下一步将MicroRAAD应用于急性HCV检测，包括： 1)开发并优化针对基因组保守区域的HCV检测新方法。对于HCV检测 设计中，我们将比较5 '非编码区（5'-NCR）和3 '非翻译区（3' UTR），这两个区域都是 用于临床HCV病毒载量测定。虽然5 '-NCR用于蛋白质免疫测定，但已知这种变异性 该区域与世界范围内流行的序列有些不同。因此，我们还将考虑以下的3 'UTR： RT-LAMP设计。我们将为每个区域产生至少两个引物，并将比较扩增时间（TTA）， 检测限（加标0-10^7 IU/mL购自ATCC的合成RNA），以及对全血和 将血浆以0- 20%的浓度加标到每份样本中。 2)扩大我们与PWUD及其服务提供者的基于社区的参与性研究，以评估以HCV为重点的 在使用点检测与艾滋病毒检测相结合。这将包括执行的可行性和可接受性 在POU进行研究，以评估PWUD检测HCV的操作、临床和市场需求， 评估PWUD中的社会文化因素和护理系统性障碍如何影响他们的HCV检测和整合 进入目前的艾滋病毒护理连续体，并评估设备原型的可用性，以确定最佳的操作协议 利益攸关方之间。 这个用于使用点HCV/HIV检测的平台可以改变HIV和HCV的检测和治疗， PWUD通过将急性感染检测带到药物滥用治疗设施，针头交换和注射药物 该计划将在2010年12月25日至26日期间在所有诊所开展，并将阐明消除护理障碍所需的针对性干预措施。 参考文献： Reipold IE，Easterbrook P，Trianni A，Panneer N，Krakower D，Oncillo S，Roberts T，米勒V，Denkinger C. 病毒血症丙型肝炎感染的优化诊断：目标产物谱的开发BMC Infect Dis. 2017 11月1日;17（增刊1）：707。doi：10.1186/s12879-017-2770-5。PMID：29143620; PMCID：PMC5688443。 Duong M，Delcher C，Freeman PR，Young AM，库珀HLF.对基于药物的HCV/HIV检测的态度 在肯塔基州乡下吸毒的人。J农村卫生。2022年1月;38（1）：93-99。doi：10.1111/jrh.12564。Epub 2021 Mar 5. PMID：33666274; PMCID：PMC8418619。

项目成果

Measurement of Protein-Protein Interaction Dynamics Using Microfluidics and Particle Diffusometry.

• DOI: 10.1021/acs.analchem.2c02570
• 发表时间: 2022-11-15
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Ma, Hui;Wereley, Steven T.;Linnes, Jacqueline C.;Kinzer-Ursem, Tamara L.
• 通讯作者: Kinzer-Ursem, Tamara L.

Thinking Beyond the Device: An Overview of Human- and Equity-Centered Approaches for Health Technology Design.

• DOI: 10.1146/annurev-bioeng-081922-024834
• 发表时间: 2023-06-08
• 期刊: Annual review of biomedical engineering
• 影响因子: 9.7

From Crisis To Crisis: Impacts Of The COVID-19 Pandemic On People Living With HIV And AIDS Service Organizations In Indiana.

从危机到危机：COVID-19 大流行对印第安纳州艾滋病毒感染者和艾滋病服务组织的影响。

• DOI: 10.21203/rs.3.rs-1003567/v1
• 发表时间: 2022
• 期刊: Research square
• 作者: MacNeill,JustinJ;Linnes,JacquelineC;Rodriguez,NataliaM
• 通讯作者: Rodriguez,NataliaM

From crisis to crisis: impacts of the COVID-19 pandemic on people living with HIV and HIV/AIDS service organizations in Indiana.

• DOI: 10.1186/s12913-022-07998-0
• 发表时间: 2022-05-09
• 期刊: BMC HEALTH SERVICES RESEARCH
• 影响因子: 2.8
• 作者: MacNeill, Justin J.;Linnes, Jacqueline C.;Hubach, Randolph D.;Rodriguez, Natalia M.
• 通讯作者: Rodriguez, Natalia M.