Automated amplification-free digital RNA detection platform for rapid and sensitive SARS-CoV-2 diagnosis
Hajime Shinoda, Tatsuya Iida, Asami Makino, Mami Yoshimura, Junichiro Ishikawa, Jun Ando, Kazue Murai, Katsumi Sugiyama, Yukiko Muramoto, Masahiro Nakano, Kotaro Kiga, Longzhu Cui, Osamu Nureki, Hiroaki Takeuchi, Takeshi Noda, Hiroshi Nishimasu, Rikiya Watanabe
Abstract
Abstract In the ongoing COVID-19 pandemic, rapid and sensitive diagnosis of viral infection is a critical deterrent to the spread of SARS-CoV-2. To this end, we developed an automated amplification-free digital RNA detection platform using CRISPR-Cas13a and microchamber device (opn-SATORI), which automatically completes a detection process from sample mixing to RNA quantification in clinical specimens within ~9 min. Using the optimal Cas13a enzyme and magnetic beads technology, opn-SATORI detected SARS-CoV-2 genomic RNA with a LoD of < 6.5 aM (3.9 copies μL −1 ), comparable to RT-qPCR. Additionally, opn-SATORI discriminated between SARS-CoV-2 variants of concern, including alpha, delta, and omicron, with 98% accuracy. Thus, opn-SATORI can serve as a rapid and convenient diagnostic platform for identifying several types of viral infections.