Reagentless detection of staphylococcal enterotoxin B via electrochemical interrogation of conformational changes.

An electrochemical biosensor for staphylococcal enterotoxin B (SEB) detection has been designed on the basis of electrochemical interrogation of conformational changes. Ferrocene-labeled hairpin probe (Fc-HP) and SEB aptamer are introduced for the construction of the platform. Without SEB, the rigid construction of DNA duplex that included SEB aptamer and Fc-HP prevented Fc getting access to the electrode surface, keeping the "eT-off" state in the detection system. In the presence of SEB, the interaction between SEB and the aptamer could trigger the disruption of DNA duplex and the restoration of hairpin structure, accompanied by the increase of Fc oxidation current. The decreasing distance between the redox probe and electrode upon the nucleic acid reconfiguration substantially increased the efficiency of eT, which resulted in the enhanced Fc signal. The proposed strategy presented a wide linear detection range from 0.005 to 100 ng mL with a detection limit down to 3 pg mL (S/N = 3). To investigate the applicability and reliability of the method in real food samples such as milk samples, we compared the results between this method and the commercial ELISA kit. The relative percentage error between the two assays ranged from -6.42% to 6.31%, indicating that there was no obvious difference between the results.