Multiplexed Airborne Virus Collection and Detection at the Point-of-Care

护理点多重空气传播病毒收集和检测

• 批准号: 10361485
• 负责人: Z. Hugh Fan
• 金额: $ 36.26万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10361485
• 关键词: 2019-nCoV; Address; Aerosols; Air; Bacteria; Biological Assay; Blood Circulation; COVID-19; COVID-19 detection; COVID-19 pandemic; Caliber; Centers for Disease Control and Prevention (U.S.); Cessation of life; Clinic; Collection; Cytolysis; DNA Primers; Detection; Devices; Disease Outbreaks; Event; FluMist; Future; General Population; Genetic; Health; Health Personnel; Healthcare; Human; Infection; International; Intervention; Laboratories; Link; Liquid substance; Mass Screening; Mediating; Methods; Paper; Particle Size; Patients; Performance; Physical condensation; Point-of-Care Systems; Policy Maker; Polymerase Chain Reaction; Preparation; Protocols documentation; Public Health; Public Hospitals; RNA; RNA amplification; Reporting; Research; Resources; Reverse Transcription; Role; Route; SARS-CoV-2 transmission; Sampling; Seasons; Sensitivity and Specificity; Stream; Students; Surveys; System; Test Result; Testing; Time; Vaccines; Virion; Virus; World Health Organization; air sampling; amplification device; aqueous; base; clinical care; co-infection; design; detection sensitivity; emerging pathogen; flu; influenzavirus; innovation; interest; isothermal amplification; multiplex detection; nanometer; nanoparticle; nanoscale; particle; pathogen; personal protective equipment; point of care; point-of-care detection; real world application; respiratory; tool; transmission process; university student; viral detection; viral transmission; water vapor

Project Summary Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the most recent emerging pathogen that causes the current pandemic of COVID-19. Although the main transmission routes for SARS-CoV-2 considered by the Centers for Disease Control and Prevention (CDC) are respiratory droplets and close contact, mounting evidences have emerged that virus transmission likely also occurs by aerosols. Similarly, influenza viruses (FluV) are transmitted via respiratory droplets, aerosols, and contact. More importantly, co-infection with FluV and SARS-CoV-2 has been observed in patients. With expected concurrent circulation of SARS-CoV-2 and FluV in the future, multiplexed collection and detection of various airborne viruses are desirable for studying transmission routes, planning and allocating resources, and mitigating the impact on public health. The collection of airborne viruses requires air samplers, ideally with two following features. One is to have the collected samples analyzed at the point-of-care (POC), rather than being sent to a lab with a delay of 1-2 days in reporting the test results. The second feature is to efficiently collect virus aerosols directly into a liquid, rather than onto a filter, for subsequent virus analysis. While liquid impingers are effective in collecting microparticles (such as bacteria), they are less so for nanoparticles such as viruses. To realize these features, we have developed (1) a Viable Virus Aerosol Sampler (VIVAS) that shows high collection efficiency of virus particles by enlarging nanometer-sized virus aerosols into micrometer-sized droplets through water vapor condensation; and (2) a Valve-enabled Lysis, paper-based RNA Enrichment, and RNA Amplification Device (VLEAD) that carries out virus lysis and washing steps without pipetting, followed by reverse transcription loop- mediated isothermal amplification (RT-LAMP) for accurate genetic identification. In this proposed research, we aim to integrate VIVAS with VLEAD for multiplexed airborne virus collection and detection at POC, assess the integrated system for collection and multiplexed detection of SARS-CoV-2 and FluV, and validate the system in real-world applications by surveying airborne viruses in a health clinic and public venues. The significance of the proposed research lies in the following aspects. First, it will lead to an innovative system that can collect and detect airborne SARS-CoV-2 and FluV at POC, which will help address a major public health concern (i.e., current COVID-19 pandemic and expected concurrent circulation with FluV in the flu seasons). Second, the integrated system will provide a critically important tool to study the role of airborne route in virus transmission. If confirmed, appropriate personal protection equipment and other remedies will be employed to reduce infections among healthcare workers and the general public. Third, the system is capable of conducting environmental surveillance via efficient collection and timely detection of these viruses at POC. It can function as a rapid mass screening method for timely results that will be valuable for local policymakers to issue prompt warning to the public and for hospitals to prepare for local outbreaks and clinical care.

项目概要 严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 是最新出现的病原体， 导致当前的 COVID-19 大流行。尽管 SARS-CoV-2 的主要传播途径被认为 疾病控制与预防中心 (CDC) 发布的报告显示，呼吸道飞沫和密切接触，越来越多 有证据表明病毒传播也可能通过气溶胶发生。同样，流感病毒（FluV） 通过呼吸道飞沫、气溶胶和接触传播。更重要的是，与流感病毒和流感病毒同时感染 已在患者中观察到 SARS-CoV-2。预计 SARS-CoV-2 和 FluV 在 未来，多种空气传播病毒的多重收集和检测对于研究传播是可取的 路线、规划和分配资源以及减轻对公共卫生的影响。 空气传播病毒的收集需要空气采样器，最好具有以下两个功能。一是要有 收集的样本在即时护理 (POC) 进行分析，而不是延迟 1-2 秒送到实验室 报告测试结果的天数。第二个特点是有效地将病毒气溶胶直接收集到液体中， 而不是过滤器上，以进行后续病毒分析。虽然液体撞击器可以有效收集 对于微粒（例如细菌），对于病毒等纳米颗粒来说，情况就不那么重要了。为了实现这些功能， 我们开发了 (1) 一种活病毒气溶胶采样器 (VIVAS)，它具有高病毒收集效率 通过水蒸气将纳米大小的病毒气溶胶放大成微米大小的液滴 缩合; (2) 阀门驱动的裂解、纸质 RNA 富集和 RNA 扩增装置 (VLEAD)，无需移液即可进行病毒裂解和洗涤步骤，然后进行逆转录环- 介导等温扩增 (RT-LAMP) 用于准确的遗传鉴定。在这项拟议的研究中，我们 旨在将 VIVAS 与 VLEAD 集成，在 POC 处进行多重空气传播病毒收集和检测，评估 用于收集和多重检测 SARS-CoV-2 和 FluV 的集成系统，并在 通过调查诊所和公共场所的空气传播病毒来实现现实世界的应用。 本研究的意义在于以下几个方面。首先，它将带来创新 该系统可以在 POC 收集和检测空气中的 SARS-CoV-2 和 FluV，这将有助于解决一个重大问题 公共卫生问题（即当前的 COVID-19 大流行以及预计流感病毒与流感病毒同时传播） 季节）。其次，集成系统将为研究机载航线的作用提供至关重要的工具 在病毒传播中。如果得到确认，将提供适当的个人防护装备和其他补救措施 用于减少医护人员和公众的感染。三、系统能力 通过在 POC 有效收集和及时检测这些病毒来进行环境监测。它 可以作为一种快速大规模筛查方法，及时获得结果，这对于当地决策者来说很有价值 向公众和医院发出及时警告，为当地疫情暴发和临床护理做好准备。