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

项目摘要 严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是最近出现的病原体， 导致了当前的COVID-19大流行。虽然SARS-CoV-2的主要传播途径被认为是 由疾病控制和预防中心（CDC）是呼吸道飞沫和密切接触，安装 有证据表明，病毒传播也可能通过气溶胶发生。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，流感病毒和 SARS-CoV-2已在患者中观察到。由于预期SARS-CoV-2和FluV在 将来，对各种空气传播病毒的多重收集和检测是研究传播的需要 路线，规划和分配资源，减轻对公共卫生的影响。 收集空气传播的病毒需要空气采样器，理想情况下具有以下两个特征。一是要有 采集的样本在床旁（POC）进行分析，而不是延迟1-2分钟发送到实验室 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， 而不是在过滤器上，用于随后的病毒分析。虽然液体撞击器有效地收集 对于微粒（如细菌），它们对纳米颗粒（如病毒）的影响较小。为了实现这些特征， 我们已经开发了（1）一个活病毒气溶胶采样器（VIVAS），它显示出高的病毒收集效率 通过水蒸气将纳米大小的病毒气溶胶放大为微米大小的液滴， （2）Valve-enabled Lysis，paper-based RNA Enrichment，and RNA Amplification Device （VLEAD），其进行病毒裂解和洗涤步骤而无需移液，随后进行逆转录环- 介导等温扩增（RT-LAMP）用于精确的遗传鉴定。在这项研究中，我们 旨在整合VIVAS和VLEAD，在POC进行多重空气传播病毒收集和检测，评估 SARS-CoV-2和FluV的采集和多重检测集成系统，并验证该系统在 通过调查健康诊所和公共场所中的空气传播病毒来实现实际应用。 本研究的意义在于以下几个方面。首先，它将导致一个创新的 该系统可以在POC收集和检测空气传播的SARS-CoV-2和FluV，这将有助于解决一个主要的 公共卫生问题（即，当前的COVID-19大流行和预期与流感病毒在流感中同时传播 季节）。第二，综合系统将提供一个至关重要的工具，研究空中路线的作用 在病毒传播中。如果得到确认，将提供适当的个人防护设备和其他补救措施。 用于减少医护人员和公众的感染。第三，系统能力 通过在POC有效收集和及时检测这些病毒进行环境监测。它 可以作为一种快速的大规模筛查方法，及时获得结果，这对当地决策者来说是有价值的， 向公众和医院迅速发出警告，以便为本地爆发和临床护理做好准备。

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.