Blue-emitting SiO2-coated Si-doped ZnSeS quantum dots conjugated aptamer-molecular beacon as an electrochemical and metal-enhanced fluorescence biosensor for SARS-CoV-2 spike protein

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) which was first reported in early January 2020, continues to devastate the worlds public health system. Herein, we report on the development of a novel metal-enhanced fluorescence (MEF) and electrochemical biosensor for SARS-CoV-2 spike (S) protein. To develop the MEF biosensor, SiO2-coated Si-doped ZnSeS quantum dots (QDs) were newly synthesized and conjugated to an aptamer-molecular beacon (Apta-MB) probe. Thereafter, cationic AuNPs, used as a localised surface plasmon resonance (LSPR) signal amplifier, were self-assembled on the QDs-Apta-MB conjugate to form a QDs-Apta-MB-AuNP probe. To develop the electrochemical biosensor, the QDs-Apta-MB assay was carried out on a carbon nanofiber-modified screen-printed carbon electrode. Cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS) were used to characterize the electrode surface whilst spectrophotometric, spectroscopic, fluorescence polarization and electron microscopic techniques were used to characterize the materials. Under optimal experimental conditions, the QDs binding to the Apta-MB, quenched the QDs' fluorescence and with SARS-CoV-2 S protein binding to the Apta-MB, LSPR signal from cationic AuNPs of different sizes and shapes were used to tune the fluorescence signal to obtain enhanced sensitivity. On the other hand, using [Fe(CN)(6)]/K3-/(4-) buffered with NaAc-KAc-TrizmaAc-KSCN-Borax as the electrolyte solution, anodic peaks of the QDs from the CV and DPV plots were unravelled. Electrochemical detection of SARS-CoV-2 S protein was accomplished by a systematic increase in the QDs anodic peak current generated from the DPV plots. The limits of detection obtained for the SARS-CoV-2 S protein were 8.9 fg/mL for the QDs-Apta-MB-AuNP MEF probe and similar to 0.5 pg/mL for the QDs-Apta-MB electrochemical probe. Detection of SARS-CoV-2 S protein in saliva was demonstrated using the QDs-Apta-MB-AuNP MEF probe.