Label-free electrochemical immunosensor for highly sensitive COVID-19 spike protein detection

The ongoing coronavirus pandemic responsible for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly increased the rate of global death and infections due to variant mutations (such as Delta and Omicron). While specifically developed and approved vaccines can limit the spread of disease in a population and severity of resulting symptoms, none have been demonstrated to effectively prevent infection altogether. Thus, reliable early diagnosis of COVID-19 is critical to identify positive cases to help contain the outbreak. Herein we report a label-free electrochemical immunosensor for rapid diagnosis of COVID-19 by using nitrogen-doped holey graphene (N-HRGO) as a nanocarrier decorated with thionine (TH) molecules as electrochemical indicators. With the spike protein located on the surface of the COVID-19 particles as the model target, the as-prepared electrochemical immunosensor could detect the presence of the COVID-19 spike protein over a wide linear range (1 pg mL^-1-10 ng mL^-1) with a low detection limit (0.3 pg mL^-1). In addition, the developed electrochemical immunosensor exhibited an excellent selectivity (with insignificant current changes towards interfering proteins comparing with COVID-19 spike protein), a good reproducibility and long-term storage stability. Importantly, the electrochemical immunosensor thus developed could successfully and reliably detect the spike protein of COVID-19 in saliva and human serum complex samples. Thus, the as-prepared label-free electrochemical immunosensor can achieve rapid and sensitive detection of the COVID-19 spike protein, as a promising clinical diagnosis tool in monitoring the progression of COVID-19.