RAPID: Collaborative Research: One-step Express Test for Presymptomatic Detection to Prevent COVID-19 Spread

严重急性呼吸综合征冠状病毒（SARS-CoV-2）的快速和失控爆发是一场前所未有的突发公共卫生事件，需要立即予以关注，以降低发病率和死亡率。在新型冠状病毒（COVID-19）爆发期间，数千名没有明显症状的潜在感染者似乎在全球大流行疾病的传播中发挥了作用，美国的感染率很高。解决这一问题需要开发新的病毒检测技术。重要的是，必须有简单、可靠和具有成本效益的方法，以便在大量感染者中，包括没有明显症状的感染者中，及早发现病毒。此外，预计这些新方法将允许在疾病暴发条件下评估大量测试样本，并成为了解免疫水平的关键。有了这个奖项，化学部门的化学测量和成像项目正在支持博士的研究。Slava V. Rotkin, Daniel J. Hayes, Suresh V. Kuchipudi（宾夕法尼亚州立大学，PSU）和Tetyana Ignatova博士（北卡罗来纳大学格林斯博罗分校，UNCG）及其合作者开发了一种新颖，准确，廉价的预筛选测试，该测试承担了检测少量活病毒颗粒的重大挑战。新的传感途径解决了在疾病早期阶段检测病毒的重要问题，以及其他方法的检测结果不太明显的问题，这些问题限制了清晰流行病学图像的创建。该项目研究了一种新的化学方法，在制造微型液滴的过程中，通过对附着在病毒外部的纳米颗粒施加磁力，以及病毒与周围环境之间的作用力，人们可以“看到”浓缩的活病毒颗粒，所有这些都是在3d打印传感器设备中进行的。这种新型传感器具有巨大的潜力，可以快速、简便地在需要的地点提供有关病毒存在的信息，以及它在病毒脱落机制和预防疾病传播方面的作用。预计这项技术可用于检测人类和动物中其他重要的基于病毒的疾病。该项目旨在开发一种用于个人护理快速病毒检测的新技术，以应对但不限于COVID-19。具体而言，该项目的目的是在发现严重症状之前，及早检测到少于1 000个病毒颗粒的病毒剂量，这相当于感染者呼吸道标本中的典型水平。由于与分析物发生化学反应的基本限制，实现基于非核酸的方法如此低的检测限是一项具有挑战性的任务。在这个项目中，通过将荧光标记的磁性纳米颗粒与SARS-CoV-2冠状病毒的糖蛋白选择性结合，然后进行水两相分离，来提高分析物的浓度。该项目旨在回答以下几个基本问题：通过血管紧张素转换酶2标记的染色标签和纳米颗粒对病毒表面的功能化来识别活病毒；在乳状液中瞬态液滴的微体积中加速这种“三明治”组装的化学和物理过程；以及化学和材料科学的途径，选择水和有机溶剂的适当组合，使乳液能够有效地两相分离病毒颗粒。该项目可能产生一种早期（症状前）检测活病毒的新技术，从而有可能在需要的地点提供预先筛选试验，以预防病毒的进一步爆发和疾病的传播。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The fast and untamed outbreak of severe acute respiratory syndrome coronavirus (SARS-CoV-2) is an unprecedented public health emergency that requires immediate attention to reduce morbidity and mortality. During the novel corona virus (COVID-19) outbreak, it appears that thousands of potentially infected individuals showing no apparent symptoms have played a role in propagating a global pandemic disease, with a high infection level in US. Addressing this issue requires development of new technologies for testing for the virus. Importantly, it is crucial to have available simple, reliable, and cost-effective methods for early detection of the virus in large numbers of infected individuals, including those who have no apparent symptoms. Furthermore, such new methods are anticipated to allow for assessment of a high volume of test samples under disease outbreak conditions and be key to understanding immunity levels. With this award, the Chemical Measurement and Imaging Program in the Division of Chemistry is supporting the research of Drs. Slava V. Rotkin, Daniel J. Hayes, Suresh V. Kuchipudi (The Pennsylvania State University, PSU), and Dr. Tetyana Ignatova (University of North Carolina at Greensboro, UNCG) and their collaborators to develop a novel, accurate, and inexpensive prescreening test that takes on the grand challenge of sensing small numbers of live virus particles. The new sensing route addresses important concerns about virus detection at early stages of the disease and less-than-obvious test results from other methods, which have limited the creation of a clear epidemiologic picture. This project investigates a new chemical approach that allows one to "see" concentrated live virus particles by using a combination of magnetic forces applied to nanoparticles bound to the outside of the virus and forces between the virus and its surroundings during creation of miniature liquid drops, all taking place inside a 3-D printed sensor device. The new sensor has great potential to deliver rapid, easy, and at the site of need, information about virus presence, as well as its role regarding mechanisms of viral shedding and preventing the spread of the disease. This technology is envisioned to be useful in detection of other important virus-based diseases, both in humans and animals.The project aims at the development of a new technology for a personal care express viral test – responding but not limited to, COVID-19. Specifically, the project is directed at enabling detection of a viral dose of less than 1,000 virus particles, which corresponds to a typical level in respiratory specimens of an infected individual well in advance of when severe symptoms can be perceived. Achieving such a low limit of detection for non-nucleic acid-based methods is a challenging task, due to fundamental limits for chemical reaction with an analyte. In this project, additional concentration of analyte is approached by combining selective binding of fluorescently tagged magnetic nanoparticles to the glycoprotein of the SARS-CoV-2 corona, followed by aqueous two-phase separation. The project is directed at answering a number of fundamental questions regarding: recognition of live viruses via functionalization of the virus surface with angiotensin converting enzyme 2 labeled staining tag and nanoparticles; chemical and physical processes of acceleration of such a "sandwich" assembly in the micro-volume of transient droplets in the emulsion; and chemical and materials science avenues for selecting a proper combination of water and organic solvents for an emulsion that allows an efficient two-phase separation of viral particles. The project may result in a new technology for early (presymptomatic) detection of the live virus, which offers the possibility of providing at the site-of-need a pre-screening test for prevention of further viral outbreaks and spread of the disease.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.