Summary CRISPR-based diagnostics (CRISPR-dx), including the Cas12-based DETECTR and Cas13-based SHERLOCK Class 2 CRISPRs, have been used to detect the presence of DNA or RNA from pathogens, such as the 2009 pandemic influenza virus A (IAV) and the 2019 novel coronavirus SARS-CoV-2. Here, we describe the collateral single-stranded DNA cleavage with Class 1 type I CRISPR-Cas3 and highlight its potential for development as a Cas3-mediated rapid (within 40 min), low-cost, instrument-free detection method for SARS-CoV-2. This assay, which we call Cas3-Operated Nucleic Acid detectioN (CONAN), not only detects SARS-CoV-2 in clinical samples, but also offers specific detection of single-base-pair mutations in IAV variants. This tool allows rapid and accurate point-of-care testing for patients with suspected SARS-CoV-2 or drug-resistant IAV infections in hospitals. Graphical abstract Highlights • Type I CRISPR-Cas3 shows trans -cleavage activity on nearby nonspecific ssDNA • We developed CONAN as a method to easily and accurately detect viral RNA • CONAN provides rapid, low-cost, and instrument-free detection of SARS-CoV-2 and IAV • CONAN also allows the specific detection of single-base-pair mutations Diagnostics; Virology
【저자키워드】 Virology, diagnostics, 【초록키워드】 SARS-CoV-2, pandemic, Mutation, Infection, Influenza virus, variants, hospitals, RNA, 2019 novel coronavirus, point-of-care, clinical samples, DNA, Patient, cleavage, Pathogens, Viral RNA, Abstract, type I, class, acid, offer, single-stranded DNA, ssDNA, IAV, highlight, detect, provide, Type, drug-resistant, nonspecific, 【제목키워드】 SARS-CoV-2, Influenza virus,