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The workflow diagram of COVID-19 MCTOP assay The COVID-19 MCTOP detection consists of four stages: Step 1 is rapid nucleic acid preparation of 15 min, the second step is RT-MCDA reaction of 40 min, and in the third step, Cas12b-gRNA complex is centrifuged into the RT-MCDA reaction mix for 15 min of target cleavage and CRISPR/Cas-based detection. Cleavage of reporter molecule (ssDNA) can be reported by real-time fluorescence (Step 3) and lateral flow biosensor of 5 min (Step 4).
