Development of a handheld rapid air sensing system to monitor and quantify SARS-CoV-2 in aerosols in real-time

项目摘要 能够快速监测蒸发成液滴的小于5 μm的气溶胶颗粒中的SARS-CoV-2 细胞核并悬浮在空气中--在出现时， 在COVID-19期间，当人们返回社区、工作场所和学校时， 流行病然而，目前基于酶的方法缺乏灵敏度、速度、简单性，并且需要实验室检测。 设备，因此缺乏实时呈现点（POP）监视的能力。在没有 尽管有实时监测持久性有机污染物的能力，但对SARS-CoV-2的传播仍然知之甚少。在本申请中， 一个多学科的研究方法，集成了快速动力学化学自动连接，非， 酶等温信号放大，固态电子学，和生物光子学提出，使 开发一种新的空气监测系统（AMS），用于检测和定量雾化的SARS-CoV-2， 实时呈现的点。病毒培养方案、空气采样技术和 单光子探测能力将为合作研究奋进提供框架， 解决COVID-19和SARS-CoV-2气溶胶传播之间知识差距的新范式 传输因此，该提案旨在改变COVID-19目前的研究方式， 提供了一个工具，使无与伦比的研究，将大大提高目前的知识基础。这些 变革性研究最终可以引导一个新的调查领域，从而更好地理解 COVID-19传播，如病毒暴露与风险、病毒衰减率与传染性、病毒载量与传染性 SARS-CoV-2空气传播的剂量。至少，拟议的三个研究目标将提供 对COVID-19气溶胶传播的基本了解。首先，当前的空气采样系统使用多级采样器。 需要几个小时才能完成的工作流程，需要实验室设备、试剂和大量的动手操作 时间目标1的目标是将联合收割机空气采样和检测结合到一个单步实时POP AMS中 在不需要实验室设备或试剂的情况下，在不到5分钟的时间内得出SARS-CoV-2定量结果的设备。 其次，吸入鼻腔和肺部的病毒接种物或初始剂量的病毒已与 与疾病的发作和严重程度有关。目标2的目标是优化和验证AMS以关联读数 用组织培养感染剂量（TCID 50）和逆转录 聚合酶链反应（RT-PCR）定量。这些参数随后可应用于人类研究 确定SARS-CoV-2气溶胶传播的人体感染剂量。第三，实地测试， 医院将提供在高风险环境中测试AMS性能的方法。目标3的目的 用RT-PCR循环阈值（Ct）值和细胞培养物TCID 50活力校准AMS测量值 结果，然后与来自世界各地的高风险环境的结果进行基准测试， SARS-CoV-2气溶胶浓度与全球感染率，作为建立阈值水平的可能性。

Project Summary The ability to rapidly monitor SARS-CoV-2 in aerosol—drop particles <5 μm in size that evaporate into droplet nuclei and become suspended in air—at the point of presentation is critical to managing the risk of infection by airborne transmission as people return to their communities, workplaces, and schools during the COVID-19 pandemic. However, current enzyme-based methods lack sensitivity, speed, simplicity, and require lab equipment—hence, lack the capability for real-time point-of-presentation (POP) monitoring. In the absence of a real-time POP monitoring capability, SARS-CoV-2 transmission remains poorly understood. In this application, a multidisciplinary research approach that integrates innovations in rapid-kinetic chemical auto-ligation, non- enzymatic isothermal signal amplification, solid-state electronics, and biophotonics is proposed to enable the development of a novel air monitoring system (AMS) that detects and quantifies aerosolized SARS-CoV-2 at the point of presentation in real-time. Recent advances in viral culturing protocols, air sampling technology, and single-photon detection capability will provide the framework for a collaborative research endeavor to establish a new paradigm to address the knowledge gap between the spread of COVID-19 and SARS-CoV-2 aerosol transmission. Therefore, the proposal is aimed at transforming the way COVID-19 is currently researched by providing a tool to enable unparalleled studies that will significantly advance the current knowledgebase. These transformative studies could ultimately guide a new field of investigations that lead to a better understanding of COVID-19 spread, such as viral exposure vs. risk, viral decay rate vs. infectivity, and viral load vs. infectious dose in SARS-CoV-2 airborne transmission. At a minimum, the proposed three research objectives will provide a basic understanding of COVID-19 aerosol transmission. Firstly, current air sampling systems use a multi-step workflow that takes several hours to complete and requires lab equipment, reagents, and significant hands-on time. The goal of objective 1 is to combine air sampling and detection into a one-step real-time POP AMS device that yields SARS-CoV-2 quantification results in less than 5 minutes, without lab equipment or reagents. Secondly, viral inoculum, or initial dose of virus, aspirated into the nasal cavity and lungs has been associated with disease onset and severity. The goal of objective 2 is to optimize and validate AMS to correlate readings from the air monitoring device with tissue-culture infectious dose (TCID50) and reverse transcription polymerase chain reaction (RT-PCR) quantities. These parameters can then later be applied to Human studies to determine the Human infectious dose of SARS-CoV-2 by aerosol transmission. Thirdly, field-based testing in hospitals will provide a means to beta test AMS performance in high-risk environments. The goal of objective 3 is to calibrate AMS measurements with RT-PCR cycle-threshold (Ct) values and cell-culture TCID50 viability results and then benchmark with results from high-risk environments taken from around the world to correlate SARS-CoV-2 aerosol concentrations with global infection rate, as a potential for establishing threshold levels.