Rapid detection of infectious viral particles by cluster induced exhaustive reaction

摘要 新冠肺炎疫情对中国的卫生、经济、环境和移民构成了巨大挑战 美国。诊断能力的增强和限制政策已广泛实施到 遏制病毒的传播。然而，美国仍有20多万人死亡，50万人住院 独自一人。这种名为严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的病毒具有很高的传染性 早期感染，尤其是年轻人，在病毒活跃传播时通常没有任何症状。 在社区中通过说话和呼吸。这两种传播途径通常会排放大量气体。 与咳嗽和打喷嚏相比，气雾剂中的病毒颗粒数量更少，因此很难检测到。尽管 可用的诊断工具种类很多，目前的现场诊断技术还不够 检测受感染者排放的气雾剂中低数量病毒颗粒的灵敏度和速度。金子 标准方法，聚合酶链式反应及其变异体，只有在实验室环境下才能极其敏感地 检测低拷贝数的病毒。排放的气溶胶是有吸引力的临床诊断目标，因为采样 是非侵入性的，检测结果直接决定了一个人是否在传播病毒。至 针对上述挑战，我们将开发一种基于聚类诱导穷举的快速诊断技术 反应(CIER)以极少的病毒敏感度和一次性装置的成本几分钱，使这种 纸质设备可以在任何地方频繁使用，以在感染患者之前立即识别该患者 将SARS-CoV-2进一步传播到社区。CIRE在理论上实现了信号的无限放大，而 保持了很高的特异性，因此非常适用于检测SARS-CoV-2颗粒中只有致密的 抗原簇可以触发CIER传感器。提出了两个具体的目标：(1)构建CIER传感器 组装抗体、间隔区和HRP部分，优化检测SARS-CoV-2的CIER设计 具有超高灵敏度的病毒颗粒，以及(2)快速定量呼吸中的SARS-CoV-2病毒颗粒 用于POC诊断的表面，并通过多重检测区分流感病毒和SARS-CoV-2。这 变革性诊断技术对SARS-CoV-2的检测将具有超高的敏感性和特异性 病毒在呼出的空气中，同时成本低，任何人都易于使用。CIER纸的使用不仅会 立即识别传播病毒的人，也节省了巨大的资源浪费，以遏制大流行 一小部分有传染性的人。

Summary The COVID-19 pandemic has posed enormous challenges to health, economy, environment and immigration in the US. Enhancement of diagnosis capacity together with restriction policies has been widely implemented to contain the virus spreading. However, there are still over 200,000 deaths and half million hospitalized in the US alone. The virus, Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2), is highly contagious while early infection particularly in young adults often exhibits no symptoms when they are actively transmitting viruses in the community through speaking and breathing. These two transmission approaches normally emit much lower number of virus particles in aerosols than coughing and sneezing, and thus are difficult to detect. Despite many types of diagnostic tools available, current on-site diagnostic technologies do not have the sufficient sensitivity and speed to detect low number of virus particles in aerosols emitted by infected people. The gold standard method, polymerase chain reaction (PCR), and its variants are only ultrasensitive in the lab setting to detect low copy number of viruses. Emitted aerosols are attractive target for clinical diagnosis because sampling is not invasive, and the detection results directly determine whether a person is spreading viruses or not. To address the above challenge, we will develop a rapid diagnostic technology based on cluster induced exhaustive reaction (CIER) with the few virus sensitivity and disposable device with the cost of pennies, so that this kind of paper device can be used frequently anywhere to identify an infectious individual immediately before he/she further spreads SARS-CoV-2 to the community. CIER enables theoretically unlimited amplification of signal while maintaining high specificity, and thus is highly applicable to detect SARS-CoV-2 particles where only the compact cluster of antigens can trigger the CIER sensor. Two specific aims are proposed: (1) Construct CIER sensor by assembling antibodies, spacer and HRP moieties, and optimize CIER’s design for detecting of SARS-CoV-2 virus particles with ultra-high sensitivity, and (2) Rapidly quantify SARS-CoV-2 virus particles in breath and on surface for POC diagnosis, and distinguish influenza virus from SARS-CoV-2 by multiplexed detection. This transformative diagnostic technology will possess ultrahigh sensitivity and specificity to detect SARS-CoV-2 viruses in exhaled air while in low cost and simple to use by any person. The use of CIER paper will not only immediately identify virus spreading people, but also save huge waste of resources to contain pandemic by a small percentage of contagious individuals.