Electrochemical pan-variant detection of SARS-CoV-2 through host cell receptor-mimicking molecular recognition

The persistent emergence of new SARS-CoV-2 variants has presented significant challenges to vaccines and antiviral therapeutics, highlighting the need for the development of methods that ensure variant-independent responses. This study introduces a unique sensor capable of electrochemically detecting SARS-CoV-2 across a wide range of variants. The comprehensive detection is achieved by using a peptide-DNA hybrid, R7-02, as the capture probe, mimicking the binding interface between a SARS-CoV-2 spike protein and a host cell receptor, hACE2. Since the first step of viral infection is the binding of the spike protein to hACE2 regardless of variant type, the hACE2-mimicking probe can naturally acquire the pan-variant recognition capability. In constructing the sensor, the R7-02 probes are positioned on electrodes via a tetrahedral DNA nanostructure for enhanced detection efficiency. Since R7-02 directly captures the externally-exposed spike protein, our approach does not require sample pretreatments, such as virus particle lysis, unlike conventional diagnostic methods. The R7-02embedded sensor demonstrated high sensitivity towards Omicron and its major subvariants-commonly known as 'stealth Omicron' (BA.5, BA.2.75, BQ.1.1, and XBB.1.5)-with a detection limit as low as 811.9 pM, along with robust specificity for SARS-CoV-2 against influenza and other human coronaviruses. The sensor also successfully detected SARS-CoV-2 directly from non-treated saliva samples of COVID-19-positive patients. Given the comprehensive and sensitive detection capability, combined with its simple operation, our receptormimicking probe-based electrochemical sensor holds the potential to be a sustainable and effective point-ofcare diagnostic tool, offering a promising solution to the constant challenges posed by the endemic presence of SARS-CoV-2.