Dual Cascade DNA Walking-Induced “super On” Photocurrent Response for Constructing a Novel Antibiotic Biosensing Method

The construction of effective methods for the convenient testing of antibiotic residues in real samples has attracted considerable interest. Herein, we designed a dual cascade DNA walking amplification strategy and combined it with the controllable photocurrent regulation of a photoelectrode to develop a novel photoelectrochemical (PEC) biosensing method for antibiotic detection. The photoelectrode was prepared through the surface modification of a glassy carbon electrode with the TiO2/CdS QDs nanocomposite synthesized by an in situ hydrothermal deposition method. The strong anodic PEC response of the nanocomposite could be well inhibited by the introduction of a silver nanoclusters (Ag NCs)-labeled DNA hairpin onto its surface. Upon the target biorecognition reaction, an Mg2+-dependent DNAzyme (MNAzyme)-driven DNA walking was triggered to release another MNAzyme strand-linked streptavidin (SA) complex. As this SA complex could serve as a four-legged DNA walker, its cascade walking on the electrode surface not only released Ag NCs but also caused the linking of Rhodamine 123 with the electrode to realize the "super on" photocurrent output. By using kanamycin as the model analyte, this method showed a very wide linear range from 10 fg mL-1 to 1 ng mL-1 and a very low detection limit of 0.53 fg mL-1. Meanwhile, the simple photoelectrode preparation and the aptamer recognitionbased autonomous DNA walking resulted in the convenient manipulation and excellent repeatability. These unique performances determine the great potential of the proposed method for practical applications.