Direct detection of SARS-CoV-2 using CRISPR-Cas13a and a mobile phone-based device. Fozouni. et al.

We report the development of an amplification-free CRISPR-Cas13a assay for direct detection of SARS-CoV-2 from nasal swab RNA that can be read with a mobile phone microscope. The assay achieved ~100 copies/uL sensitivity in under 30 minutes of measurement time and accurately detected pre-extracted RNA from a set of positive clinical samples in under 5 minutes. We combined crRNAs targeting SARS-CoV-2 RNA to improve sensitivity and specificity, and directly quantified viral load using enzyme kinetics. Integrated with a reader device based on a mobile phone, this assay has the potential to enable rapid, low-cost, point-of-care screening for SARS-CoV-2.

本报告阐述了无需扩增的CRISPR-Cas13a检测方法的发展，该方法可直接从鼻拭子RNA中检测SARS-CoV-2，并可利用手机显微镜进行读取。该检测方法在30分钟内即可实现约100拷贝/微升的灵敏度，并在5分钟内准确检测到一组阳性临床样本中预先提取的RNA。我们结合了针对SARS-CoV-2 RNA的crRNA以提高灵敏度和特异性，并利用酶动力学直接量化病毒载量。该检测方法与基于手机的读取设备集成，有望实现快速、低成本、床旁的SARS-CoV-2筛查。