Amplification-free RNA detection with CRISPR-Cas13

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 CRISPR-based amplification-free digital RNA detection (SATORI), by combining CRISPR-Cas13-based RNA detection and microchamber-array technologies. SATORI detected single-stranded RNA targets with maximal sensitivity of similar to 10 fM in <5min, 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 similar to 5 fM levels. Therefore, we hope SATORI will serve as a powerful class of accurate and rapid diagnostics.

Automated summary

The study introduces a novel CRISPR-based platform called SATORI, which allows amplification-free digital RNA detection with high sensitivity and specificity in a short period of time. The simultaneous use of multiple guide RNAs enhances sensitivity, enabling the detection of SARS-CoV-2 RNA at low levels.