Journal
COMMUNICATIONS BIOLOGY
Volume 4, Issue 1, Pages -Publisher
NATURE PORTFOLIO
DOI: 10.1038/s42003-021-02001-8
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Funding
- JST CREST [JPMJCR19H5]
- AMED [20he0622032h0001, 19fk0108113, 20fk0108270h0001]
- Mitsubishi Foundation Grant for Academic Research on Infectious Diseases
- JSPS [20H05931]
- Institute of Medical Science
- University of Tokyo
- Joint Usage/Research Center program of Institute for Frontier Life and Medical Sciences Kyoto University
- Grants-in-Aid for Scientific Research [20H05931] Funding Source: KAKEN
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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.
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.
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