CRISPR-based nucleic-acid detection is an emerging technology for molecular diagnostics. However, these methods generally require several hours and could cause amplification errors, due to the pre-amplification of target nucleic acids to enhance the detection sensitivity. Here, we developed a platform that allows “CRI S PR-based a mplifica t i o n-free digital R NA detect i on (SATORI)”, by combining CRISPR-Cas13-based RNA detection and microchamber-array technologies. SATORI detected single-stranded RNA targets with maximal sensitivity of ~10 fM in <5 min, with high specificity. Furthermore, the simultaneous use of multiple different guide RNAs enhanced the sensitivity, thereby enabling the detection of the SARS-CoV-2 N-gene RNA at ~5 fM levels. Therefore, we hope SATORI will serve as a powerful class of accurate and rapid diagnostics. The authors develop a platform (SATORI) that enables accurate and rapid detection of single-stranded RNA at a single-molecule level without a pre-amplification step. As a proof-of-concept, they demonstrate its utility in detecting the SARS-CoV-2 N gene at a minimum concentration of ~5 fM, which is much lower than other amplification-free CRISPR-Cas-based methods.
【저자키워드】 Nanobiotechnology, Enzymes, 【초록키워드】 diagnostics, RNA, molecular diagnostics, CRISPR, amplification, nucleic acid, sensitivity, specificity, nucleic acids, target, molecular, N gene, utility, platform, Detection sensitivity, guide RNAs, Concentration, proof, N-gene, SARS-CoV-2 N gene, single-stranded, guide RNA, ENhance, detect, develop, much lower, the SARS-CoV-2, 【제목키워드】 RNA,