Sample-to-Answer, Rapid, Multiplexed and PCR-Free Diagnostics of Arboviral Diseases in Resource Limited Settings

在资源有限的情况下对虫媒病毒疾病进行从样本到答案、快速、多重且无 PCR 的诊断

• 批准号: 10528470
• 负责人: Ahmet Ali Yanik
• 金额: $ 75万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-23 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10528470
• 关键词: Acute; Animals; Antibodies; Antibody Formation; Arbovirus Infections; Arboviruses; Archives; Area; Biological Assay; Blood; Blood specimen; California; Chikungunya virus; Chronic; Clinical; Collaborations; Country; Dengue Virus; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease Outbreaks; Disease Outcome; Disease Surveillance; Early Diagnosis; Education; Endemic Diseases; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemic; Fever; Foundations; Geography; Goals; Grant; Grenada; Health; Human; Industry; Infection; Infectious Disease Epidemiology; Infrastructure; Island; Kenya; Laboratories; Location; Measurable; Measurement; Medical; Microfluidics; Molecular; Molecular Virology; National Institute of Allergy and Infectious Disease; Nucleic Acid Amplification Tests; Patients; Phase; Plasma; Population; Populations at Risk; Proteins; Public Health; Recombinants; Research; Resource-limited setting; Resources; Risk; Sampling; Scheme; Serum; Special Equipment; Surveillance Program; Symptoms; System; Technology; Testing; Time; Training; United States National Institutes of Health; Universities; Validation; Viral; Viral Antigens; Virus Diseases; Work; Zika Virus; anti-IgM; arboviral disease; arthropod-borne; biodefense; clinical diagnostics; cost; cost effective; design; detection assay; diagnostic platform; diagnostic strategy; diagnostic technologies; diagnostic tool; disease diagnostic; disease transmission; emerging pathogen; experimental study; field study; high risk; human disease; improved; multidisciplinary; multiplex detection; nanoengineering; nanophotonic; new technology; novel; novel diagnostics; novel strategies; pathogen; point of care; portability; programs; prototype; tool; transmission process; viral detection

Arthropod-borne viruses (arboviruses) comprise many of the most important ‘emerging pathogens’ due to their geographic spread and their increasing impact on vulnerable human populations. There is urgent need for easy-to- operate and rapidly deployable diagnostic tools that can handle blood samples in a closed sample-to-answer manner. Here, we propose to develop a novel diagnostic technology that can detect viral antigens in an inexpensive, ultrasensitive, specific, and multiplexed manner. We will develop our novel approach into standalone tool with a detection capability at attomolar sensitivities (comparable to nucleic acid amplification tests) to diagnose arboviral infections with minimal user interference. The integrated diagnostic platform will utilize a novel surrogate approach, microfluidic integration, and a multiplexed detection scheme with the capacity to distinguish arboviral infections. The system will be designed to initiate diagnosis from serum/plasma/blood and provide a sample-to-answer diagnostic within less than 35 minutes using less than 100 µL blood samples at a cost of $2 per test. Collaborative work proposed for this NIH/NIAID R01 Grant involves integration of nanophotonic engineering (Yanik Group), molecular virology (Pinsky Group), and infectious diseases epidemiology (LaBeaud Group) to build and field-test our novel point-of-care viral diagnostic platform with Windward Islands Research and Education Foundation (WINDREF) and St. George’s University teams (Macpherson, Waechter and Noel Groups). Preliminary validation tests with patient samples will be initially performed at Stanford Medical Facility in collaboration with LaBeaud and Pinsky groups. Subsequently, three prototypes will be transferred to Grenada for field-testing initially at central laboratories then to resource-poor settings in small towns. Yanik group will provide the necessary expertise for integration of molecular and nanoengineering components and demonstration of a practical prototype as well as evaluating the application of prototype(s) developed under this proposal with patient samples (LaBeaud and Pinsky Groups). System will be iteratively optimized and a rugged platform suitable for field settings will be developed.

节肢动物传播的病毒（虫媒病毒）由于其地理分布及其对脆弱人群的影响越来越大，构成了许多最重要的“新兴病原体”，因此迫切需要易于操作且可快速部署的能够处理血液的诊断工具。在这里，我们建议开发一种新型诊断技术，能够以廉价、超灵敏、特异性和多重方式检测病毒抗原，我们将把我们的新方法开发成具有检测功能的独立工具。能力在该集成诊断平台将利用一种新颖的替代方法、微流体集成和能够区分虫媒病毒感染的多重检测方案，以阿托摩尔灵敏度（与核酸扩增测试相当）来诊断虫媒病毒感染。旨在从血清/血浆/血液开始诊断，并在 35 分钟内使用少于 100 µL 的血液样本提供样本到答案诊断每次测试费用为 2 美元，NIH/NIAID R01 资助计划的合作工作涉及纳米光子工程（Yanik 小组）、分子病毒学（Pinsky 小组）和传染病流行病学（LaBeaud 小组）的整合，以构建和现场测试我们的小说。与向风群岛研究和教育基金会 (WINDREF) 和圣乔治大学团队（Macpherson、Waechter 和 Noel Groups）合作的即时病毒诊断平台。最初将与 LaBeaud 和 Pinsky 团队合作，在斯坦福医疗机构对患者样本进行初步验证测试，随后，三个原型将被转移到格林纳达，首先在中心实验室进行现场测试，然后再转移到资源匮乏的小镇雅尼克。小组将提供必要的专业知识，以整合分子和纳米工程组件，演示实用原型，并评估根据本提案开发的原型与患者样本的应用（LaBeaud 和 Pinsky）组）。系统将被迭代优化，并开发出适合现场设置的坚固平台。