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，Luminostics将利用我们的去风险和可扩展的CLIP平台 开发一种基于智能手机的快速护理点（POC）诊断方法，用于检测SARS-CoV-2抗原 呼吸道标本Luminostics的CLIP技术可实现高灵敏度的侧流免疫分析 （LFA）使用专有的“纳米荧光粉”，结合消费者智能手机光学和先进的信号 用于读出的处理算法。由于迫切需要经过验证的快速POC检测工具， 活跃的COVID-19疾病，该项目将迅速采用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一个完整的结果。 结果，增加了继续传播的可能性。此外，大多数NAAT必须在 实验室设置使用昂贵的设备。FDA紧急使用下批准的POC测试 COVID-19的授权（EUA）也需要购买专用设备，限制了广泛使用。 因此，该项目将开发、验证并获得FDA EUA用于CLIP-COVID，这是一种基于智能手机的快速检测 用于基于SARS-CoV-2的超灵敏免疫测定检测活动性COVID-19感染 呼吸道样本中的抗原，包括残留病毒转运培养基（VTM）、鼻咽/鼻拭子， 鼻吸出物唾液和痰当用作LFA中的标签时，Luminostics的专利持久发光 无机纳米荧光粉能够实现数量级的较低检测限（LOD）， CLIP平台上的临床灵敏度与传统的视觉读取LFA相比，仅使用智能手机 光学器件，搭配廉价的适配器和移动的应用程序，实现清晰的读数。该检测将使用高- SARS-CoV-2抗原特异性亲和单克隆抗体（mAb）作为诊断试剂。夹子 一个平台先前已经优化，以检测细菌和病毒病原体已经证实-为我们的衣原体 测试-临床灵敏度约为90%，特异性为99%，室温稳定性>15个月，使 检测试剂盒的运输、储存和储存方便。该项目旨在开发、验证和获得FDA EUA用于CLIP-COVID诊断测试，目的如下：1）筛选大量mAb， 特异性结合SARS-CoV-2抗原并选择2-3个优选的夹心mAb对; 2）开发 和CLIP-COVID组件的优化，包括测定设计、缓冲液/缀合物化学和其他 辅助组件;和3）分析/临床验证研究、生产转移和EUA提交。 该项目的预期成果是为POC COVID-19提供负担得起、货架稳定、快速、用户友好的测试 在项目启动后6个月内完成FDA EUA提交的检测。

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.