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

SARS-CoV-2 和病毒相关外泌体的微流体分离和表征

• 批准号: 10321009
• 负责人: Genevieve Marie Boland
• 金额: $ 99.91万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-21 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10321009
• 关键词: 2019-nCoV; Address; Area; Arts; Authorization documentation; Benchmarking; Biological; Biological Assay; Blood; Blood Preservation; Blood specimen; CLIA certified; COVID-19; COVID-19 detection; COVID-19 diagnosis; COVID-19 diagnostic; COVID-19 early detection; COVID-19 patient; COVID-19 point of care; COVID-19 testing; Cancer Center; Cancer Patient; Cells; Clinic; Clinical; Communities; Complex; Convalescence; Data; Devices; Diagnosis; Diagnostic; Event; Feces; Feedback; Goals; Immune; Immunology; Infection; Knowledge; Laboratories; Lead; Liquid substance; Malignant Neoplasms; Microfluidic Microchips; Microfluidics; Neoplasm Circulating Cells; Organ; Outcome; Pathway interactions; Patient Care; Patients; Phase; Plasma; Population; Production; Publishing; RADx Radical; RNA; Radiologic Health; Resolution; Resources; Running; SARS-CoV-2 infection; Saliva; Sampling; Sensitivity and Specificity; Specialist; Specimen; Specimen Handling; Technology; Testing; Time; Training; Translating; Validation; Vesicle; Viral; Virion; Virus; Whole Blood; Work; acute infection; assay development; base; cancer cell; clinical biomarkers; coronavirus disease; cost; cross reactivity; detection assay; detection limit; detection sensitivity; disease phenotype; exosome; in-vitro diagnostics; innovation; liquid biopsy; microfluidic technology; molecular diagnostics; nanoscale; new technology; novel; novel strategies; particle; preservation; research clinical testing; technology development; viral detection

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.

项目概要 由于我们缺乏超强、高效和可靠的 SARS-CoV-2 检测，因此极具挑战性。 敏感的检测方法和不断发展的病毒知识。基于标准 PCR 的检测结果仍然非常高 感染最初几天的假阴性率。临床样本的微流控处理成本低， 显示出将大多数液体活检测定转化为临床的巨大前景。我们的实验室是第一个 应用微流体技术分离血液中的循环肿瘤细胞和外泌体 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资源 用于病毒检测以及其他可能告知我们感染情况的临床生物标志物。因此，对于这支U18来说， 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， 测试其对每种生物流体的效用。为了完成这项工作，我们将完成完整的临床验证和 测定的基准测试。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 检测测定以确定检测灵敏度和特异性。此外，我们计划展示 我们的微流体装置提高了灵敏度和特异性，将导致更早检测到 SARS-CoV-2，减少此测试窗口中的假阴性。完成这项工作后，我们将拥有 收集了数据训练集并提交了完整的 EUA 计划，该计划将使 FDA 能够授权我们 用于临床的测试。