Abstract
The main protease (Mpro), which is highly conserved and plays a critical role in the replication of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), is a natural biomarker for SARS‐CoV‐2. Accurate assessment of the Mpro activity is crucial for the detection of SARS‐CoV‐2. Herein, we report a nanopore‐based sensing strategy that uses an enzyme‐catalyzed cleavage reaction of a peptide substrate to measure the Mpro activity. The peptide was specifically cleaved by the Mpro, thereby releasing the output products that, when translocated through aerolysin, quantitatively produced the signature current events. The proposed method exhibited high sensitivity, allowing the detection of Mpro concentrations as low as 1 nM without the use of any signal amplification techniques. This simple, convenient, and label‐free nanopore assay may expand the diagnostic tools for viruses.
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