Institute for Basic Science (IBS), Center for Nanomedicine, Seoul, South Korea; Center for Systems Biology, Massachusetts General Hospital Research Institute, Boston, MA, USA.
Center for Systems Biology, Massachusetts General Hospital Research Institute, Boston, MA, USA; Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, MA, USA.
Biosens Bioelectron. 2021 Apr 15;178:113049. doi: 10.1016/j.bios.2021.113049. Epub 2021 Jan 30.
Prompt diagnosis, patient isolation, and contact tracing are key measures to contain the coronavirus disease 2019 (COVID-19). Molecular tests are the current gold standard for COVID-19 detection, but are carried out at central laboratories, delaying treatment and control decisions. Here we describe a portable assay system for rapid, onsite COVID-19 diagnosis. Termed CODA (CRISPR Optical Detection of Anisotropy), the method combined isothermal nucleic acid amplification, activation of CRISPR/Cas12a, and signal generation in a single assay, eliminating extra manual steps. Importantly, signal detection was based on the ratiometric measurement of fluorescent anisotropy, which allowed CODA to achieve a high signal-to-noise ratio. For point-of-care operation, we built a compact, standalone CODA device integrating optoelectronics, an embedded heater, and a microcontroller for data processing. The developed system completed SARS-CoV-2 RNA detection within 20 min of sample loading; the limit of detection reached 3 copy/μL. When applied to clinical samples (10 confirmed COVID-19 patients; 10 controls), the rapid CODA test accurately classified COVID-19 status, in concordance with gold-standard clinical diagnostics.
快速诊断、患者隔离和接触者追踪是控制 2019 年冠状病毒病（COVID-19）的关键措施。分子检测是目前 COVID-19 检测的金标准，但在中央实验室进行，延迟了治疗和控制决策。在这里，我们描述了一种用于快速现场 COVID-19 诊断的便携式检测系统。该方法称为 CODA（CRISPR 各向异性光检测），将等温核酸扩增、CRISPR/Cas12a 的激活和单一检测中的信号生成相结合，消除了额外的手动步骤。重要的是，信号检测基于荧光各向异性的比测量，这使得 CODA 能够实现高信噪比。为了实现床边操作，我们构建了一个紧凑的独立 CODA 设备，集成了光电、嵌入式加热器和用于数据处理的微控制器。该系统在 20 分钟内完成了 SARS-CoV-2 RNA 的检测；检测限达到 3 拷贝/μL。当应用于临床样本（10 名确诊 COVID-19 患者；10 名对照）时，快速 CODA 测试准确地对 COVID-19 状态进行分类，与金标准临床诊断一致。
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