Exosome-based Non-traditional Technologies Towards Multi-Parametric andIntegrated Approaches for SARS-CoV-2

摘要 本附录是对编号PA-20-272的资金机会公告的回应 对现有NIH补助金和合作协议的行政补充。这个 补充申请是对RADx-RAD关于机会的公告的回应 额外的资源和资金支持，以促进商业化的道路。这个 机会由RADx-RAD数据协调中心(DCC)宣布并转发 通过美国国立卫生研究院种子办公室。 遏制SARS-CoV-2传播的迫切需要要求提供以下诊断方法 比定量聚合酶链式反应和抗体检测更快速、准确、灵敏和负担得起。当前的qPCR检测 是具体和敏感的，但漫长的周转时间限制了对疾病传播的干预。 抗体检测速度更快，但假阳性/阴性率很高。在此，我们建议 重新利用胞外囊泡(EV)分离和表征技术 低成本、准确、灵敏、快速(5分钟内)的SARS-CoV-2自动化检测平台， 也很实用。该设备已经过优化，可以分析血液、唾液和鼻咽(NP)。 棉签。然而，在这项提案中，我们将重点关注唾液或流口水样本(25-50微升) 可在家中轻松自取设置。样品是用自动化设备处理的 与Sognef Inc.共同开发，数据通过传输到Sognef服务器进行分析 一款可以下载到任何智能设备的应用程序。这项技术与雅培最近的技术类似 推出了新冠肺炎银卡，但具有多参数检测的优势 病毒核糖核酸(VRNA)，包括Spike(S)和核衣壳(N)蛋白编码区，揭示 即使在病毒载量低于可检测到的限度时也会感染。这种方法将减少虚假- 阳性/阴性，抗原/抗体检测的主要限制。

Abstract This supplement is in response to Funding Opportunity Announcement Number PA-20-272 Administrative Supplements to Existing NIH Grants and Cooperative Agreements. The supplement application is in response to the RADx-rad announcement on the opportunity of additional resources and funding support to facilitate the pathway to commercialization. The opportunity was announced by the RADx-rad Data Coordination Center (DCC) and relayed through the NIH SEED office. The urgent need to curb the spread of SARS-CoV-2 demands availability of diagnostics that are more rapid, accurate, sensitive and affordable than qPCR and antibody tests. Current qPCR tests are specific and sensitive, but lengthy turnaround times limit interventions against disease spread. Antibody testing is faster, but false-positive/negative rates are high. Here, we propose to repurpose extracellular vesicle (EV) separation and characterization technologies into a fully automated SARS-CoV-2 testing platform that is low-cost, accurate, sensitive, rapid (within 5 min), and practical. The device has been optimized to analyze blood, saliva, and nasopharyngeal (NP) swabs. However, in this proposal we will be focusing on saliva or drooling samples (25-50 ul) that can be easily self-obtained at-home setting. The sample is processed with an automated device developed with Sognef Inc., and data are transferred to the Sognef servers for analysis through an app that can be downloaded to any smart device. This technology is similar to Abbott's recently launched BinaxNOW COVID-19 Ag Card, but has the advantage of multiparametric detection of viral RNA (vRNA), including Spike (S) and Nucleocapsid (N) protein-coding regions that reveals infection even when viral loads are below detectable limits. This approach will decrease false- positives/negatives, the major limitation of antigen/antibody tests.