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

摘要 本补充材料是对融资机会公告编号PA-20-272的回应 对现有NIH赠款和合作协议的行政补充。的 补充申请是为了响应RADx-rad宣布的机会， 增加资源和资金支持，以促进商业化。的 这个机会是由RADx-rad数据协调中心（DCC）宣布的， 通过NIH种子办公室。 遏制SARS-CoV-2传播的迫切需要要求提供 比qPCR和抗体检测更快速、准确、灵敏和经济。当前qPCR检测 具有特异性和敏感性，但漫长的周转时间限制了针对疾病传播的干预措施。 抗体检测更快，但假阳性/阴性率很高。在此，我们建议 重新利用细胞外囊泡（EV）分离和表征技术， SARS-CoV-2自动化检测平台，成本低，准确，灵敏，快速（5分钟内）， 也很实用该设备已优化分析血液，唾液和鼻咽（NP） 棉签然而，在本提案中，我们将重点关注唾液或唾液样本（25-50 ul）， 可以很容易地在家里自己获得设置。样品用自动化设备处理 与Sognef Inc.合作开发，和数据传输到Sognef服务器进行分析， 一个可以下载到任何智能设备上的应用程序。这项技术类似于雅培公司最近在 推出BinaxNOW COVID-19 Ag Card，但具有多参数检测的优势， 病毒RNA（vRNA），包括刺突（S）和核衣壳（N）蛋白编码区， 即使病毒载量低于可检测的限度，也会感染。这种方法会减少错误- 阳性/阴性，抗原/抗体测试的主要限制。

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.