A smartphone-based rapid point-of-care test for SARS-CoV-2 detection from respiratory samples

摘要 在这个紧急补充项目中，Lighostics将利用我们的低风险和可扩展的CLIP平台 建立一种基于智能手机的快速检测SARS-CoV-2抗原的POC诊断方法 从呼吸道样本中提取。Lighostics的夹子技术使高灵敏度的横向流动免疫分析成为可能 (LFA)结合消费类智能手机的光学技术和先进的信号技术，使用专有的“纳米荧光粉” 用于读出的处理算法。因为迫切需要用于快速PoC检测的有效工具 活跃的新冠肺炎病，本项目将尽快采用CLIP技术检测SARS-CoV-2 抗原。虽然核酸扩增测试(NAAT)具有很高的准确性，但它们可能需要几天时间才能返回 结果，增加了继续传播的可能性。此外，大多数NAAT必须在 实验室环境使用昂贵的设备。根据FDA的紧急使用批准的POC测试 新冠肺炎的授权还需要购买专门的设备，这限制了广泛使用。 因此，该项目将开发、验证并获得FDA对CLIP-COVID的EUA，这是一种基于智能手机的快速测试 基于SARS-CoV-2超灵敏免疫分析检测新冠肺炎活动性感染 呼吸道样本中的抗原，包括残留病毒传输介质(VTM)、鼻咽/鼻拭子、 鼻腔吸出物、唾液和痰。当用作LFA的标签时，Lignostics的专利持久发光 无机纳米磷使检测下限(LOD)降低了数量级--因此更高 临床敏感性-在CLIP平台上，与传统的视觉阅读LFA相比，仅使用智能手机的 光学元件，配上便宜的适配器和移动应用程序，可以清晰地读出数据。化验将使用高- 抗SARS-CoV-2抗原的亲和单抗(MAbs)作为诊断试剂。这个片段 平台之前已经过优化，可以检测细菌和病毒病原体的回归-对于我们的衣原体 测试-临床灵敏度为~90%，特异度为99%，室温稳定性为&gt；15个月，使 测试套件的运输、存储和储存都很方便。该项目寻求开发、验证和获得FDA 用于CLIP-COVID诊断试验的EUA通过以下目的：1)筛选大量单抗，用于 SARS-CoV-2抗原特异性结合及2-3对首选夹心单抗的筛选；2)发展 和优化Clip-CoVID组分，包括检测设计、缓冲/结合化学等 辅助组件；以及3)分析/临床验证研究、制造转移和EUA提交。 此项目的预期结果是对PoC新冠肺炎进行负担得起、货架稳定、快速、用户友好的测试 FDA EUA提交的检测在项目启动后6个月内完成。

Abstract In this Emergency Supplemental project, Luminostics will leverage our de-risked and scale-ready CLIP platform to develop a rapid smartphone-based point-of-care (POC) diagnostic for the detection of SARS-CoV-2 antigens from respiratory specimens. Luminostics' CLIP technology enables high-sensitivity lateral flow immunoassays (LFAs) using proprietary “nanophosphors” in combination with consumer smartphone optics and advanced signal processing algorithms for readout. As there is an immediate need for validated tools for rapid POC detection of active COVID-19 disease, this project will expeditiously adapt CLIP technology for detection of SARS-CoV-2 antigens. While nucleic acid amplification tests (NAATs) have high accuracy, they can take days to return a result, increasing the potential for continued transmission. In addition, most NAATs must be processed in a laboratory setting using expensive equipment. POC tests approved under the FDA's emergency use authorization (EUA) for COVID-19 also require the purchase of specialized equipment, limiting widespread use. Thus, this project will develop, validate, and obtain FDA EUA for CLIP-COVID, a rapid smartphone-based test for the detection of active COVID-19 infection based on an ultrasensitive immunoassay for SARS-CoV-2 antigens in respiratory samples, including remnant viral transport media (VTM), nasopharyngeal/nasal swabs, nasal aspirate, saliva, and sputum. When used as labels in LFAs, Luminostics' patented persistent luminescent inorganic nanophosphors enable orders-of-magnitude lower limits of detection (LODs)—and therefore higher clinical sensitivities—on the CLIP platform compared to traditional visually-read LFAs, using only a smartphone's optics, paired with an inexpensive adapter and mobile app, for unambiguous readout. The assay will use high- affinity monoclonal antibodies (mAbs) specific to SARS-CoV-2 antigens as diagnostic reagents. The CLIP platform has been previously optimized to detect bacterial and viral pathogens has returned—for our Chlamydia test—a clinical sensitivity of ~90%, specificity of 99%, and room temperature-stability for >15 months, enabling the easy transport, storage, and stockpiling of test kits. This project seeks to develop, validate, and obtain FDA EUA for a CLIP-COVID diagnostic test through the following aims: 1) Screening of a large number of mAbs for specific binding to SARS-CoV-2 antigens and selection of 2–3 preferred sandwich mAb pairs; 2) Development and optimization of CLIP-COVID components, including assay design, buffer/conjugate chemistry, and other ancillary components; and 3) Analytical/clinical validation studies, manufacturing transfer, and EUA submission. The expected outcome of this project is an affordable, shelf-stable, rapid, user-friendly test for POC COVID-19 detection with FDA EUA submission completed within 6 months of project initiation.