Microfluidic Isolation and Characterization of SARS-CoV-2 and Virus Related Exosomes

项目摘要 由于我们缺乏超常规的技术，对SARS-CoV-2进行稳健、高效和可靠的检测是非常具有挑战性的。 灵敏的检测和不断发展的病毒知识。基于标准PCR的测定仍然导致非常高的 感染初期的假阴性率临床样品的微流体处理成本低， 显示出将大多数液体活检测定转化为临床的巨大前景。我们的实验室是最早 应用微流体技术分离血液中的循环肿瘤细胞和外来体， 癌症患者。对于SARS-CoV-2患者来说，唾液、粪便和血浆都被认为是潜在的资源 用于病毒检测以及其他可能告知我们感染的临床生物标志物。因此，对于U18， 我们将优化我们的外泌体捕获技术EVHB-Chip，用于分离完整的SARS-CoV-2病毒， 测试它对每种生物液体的效用为了完成这项工作，我们将完成完整的临床验证， 测定的基准。一旦完全优化，我们的检测方法将与现有的EUA进行比较 SARS-CoV-2检测试验，以确定检测灵敏度和特异性。此外，我们计划展示 我们的微流控装置提高了灵敏度和特异性， SARS-CoV-2，减少该测试窗口中的假阴性。在这项工作完成后，我们将有 收集了数据训练集，并提交了一份完整的EUA计划，该计划将使FDA能够授权我们的 测试将用于临床。

PROJECT SUMMARY Robust, efficient and reliable testing for SARS-CoV-2 is extraordinarily challenging due to our lack of ultra- sensitive assays and ever evolving knowledge of the virus. Standard PCR based assays still result in very high false negative rates in the earliest days of infection. Microfluidic processing of clinical samples is low cost and shows great promise for translating most liquid biopsy assay to the clinic. Our laboratory was one of the first to apply microfluidic technologies for the isolation of both circulating tumor cells and exosomes in the blood of patients with cancer. For SARS-CoV-2 patients, saliva, stool, and plasma are all thought to be potential resources for both virus detection as well as other clinical biomarkers that might inform us of infection. Thus, for this U18, we will optimize our exosome capture technology, the EVHB-Chip, for the isolation of intact SARS-CoV-2 virus, testing its utility for each of these biofluids. To complete this work, we will complete a full clinical validation and benchmarking of the assay. Once fully optimized, our detection assay will be compared against existing EUA SARS-CoV-2 detection assay to determine detection sensitivity and specificity. Further, we plan to demonstrate that the increased sensitivity and specificity enabled by our microfluidic device will result in earlier detection of SARS-CoV-2, reducing false negatives in this testing window. At the completion of this work, we will have collected the data training sets and submitted a full EUA plan that would enable the FDA’s authorization of our test to be used in the clinic.