加州大学旧金山分校Melanie Ott、Daniel A. Fletcher等研究人员，合作利用CRISPR-Cas13a和手机显微镜实现对SARS-CoV-2的无扩增检测。该项研究成果于2020年12月4日在线发表在《细胞》杂志上。

研究人员报告了一种免扩增的CRISPR-Cas13a检测技术，该技术可直接从鼻拭子RNA中直接检测SARS-CoV-2，并可通过手机显微镜对其进行读取。该测定法在30分钟的测量时间内即可达到约100拷贝每微升的灵敏度，并能在5分钟内准确地检测出一组阳性临床样品中的预提取RNA。

研究人员结合靶向SARS-CoV-2 RNA的crRNA来提高了敏感性和特异性，并使用酶动力学直接定量病毒载量。与基于手机的阅读器设备集成后，该检测方法有可能实现SARS-CoV-2的快速、低成本、即时检查。

研究人员表示，新型呼吸道病毒SARS-CoV-2已成为全球性的大流行病，但检测有症状、无症状和症状前携带者仍是难题。CRISPR诊断程序可以快速、便携和准确地进行，因此可以增强金标准的PCR测试方法。

附：英文原文

Title: Amplification-free detection of SARS-CoV-2 with CRISPR-Cas13a and mobile phone microscopy

Author: Parinaz Fozouni, Sungmin Son, María Díaz de León Derby, Gavin J. Knott, Carley N. Gray, Michael V. D’Ambrosio, Chunyu Zhao, Neil A. Switz, G. Renuka Kumar, Stephanie I. Stephens, Daniela Boehm, Chia-Lin Tsou, Jeffrey Shu, Abdul Bhuiya, Max Armstrong, Andrew R. Harris, Pei-Yi Chen, Jeannette M. Osterloh, Anke Meyer-Franke, Bastian Joehnk, Keith Walcott, Anita Sil, Charles Langelier, Katherine S. Pollard, Emily D. Crawford, Andreas S. Puschnik, Maira Phelps, Amy Kistler, Joseph L. DeRisi, Jennifer A. Doudna, Daniel A. Fletcher, Melanie Ott

Issue&Volume: 2020-12-04

Abstract: The December 2019 outbreak of a novel respiratory virus, SARS-CoV-2, has become an ongoing global pandemic due in part to the challenge of identifying symptomatic, asymptomatic and pre-symptomatic carriers of the virus. CRISPR diagnostics can augment gold-standard PCR-based testing if they can be made rapid, portable and accurate. Here 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/μL 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.

DOI: 10.1016/j.cell.2020.12.001

Source: https://www.cell.com/cell/fulltext/S0092-8674(20)31623-8