Detection of staphylococcal enterotoxin B (SEB) as a bacterial toxin causing severe food poisoning is of great importance. Herein, we developed an electrochemical aptasensor for SEB detection using a screen printed electrode modified with reduced graphene oxide (rGO) and gold nano-urchins (AuNUs). Afterward, the single-stranded DNA probe was attached to the surface of AuNUs on the modified electrode and then the specific aptamer was attached to the probe. In the presence of SEB molecules, the aptamer detached from the electrode surface and after applying the electrochemical signal generator, hematoxylin and the peak current of differential pulse voltammetry (DPV) were recorded. Due to the intercalation mechanism of hematoxylin-DNA interaction, the detachment of aptamer from electrode surface decreased the DPV peak current and the calibration graph (peak current vs SEB concentration) can be used for quantification of SEB. The cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) and also field emission scanning electron microscope imaging were used for electrode characterization. Selectivity experiments of the developed aptasensor showed a very distinct difference between SEB and other nonspecific molecules. A wide linear range from 5.0 to 500.0 fM was achieved and the detection limit was calculated as 0.21 fM. The performance of the aptasensor was checked in spiked food samples as simulated real samples and the results showed no significant difference compared to the synthetic samples. Results of selectivity and repeatability of the aptasensor were satisfactory. In addition, better recovery percentages and also lower standard deviation of aptasensor compared to a commercial ELISA kit of SEB detection proved the superior performance of the proposed aptasensor.
Keywords: Aptasensor; Electrochemical biosensor; Gold nano-urchins; Reduced graphene oxide; Staphylococcal enterotoxin B.
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