A fluorescent aptasensor for ochratoxin A detection based on enzymatically generated copper nanoparticles with a polythymine scaffold.

A fluorescence enhancement method is presented for the determination of ochratoxin A (OTA). The interaction of OTA with its aptamer causes structural changes which, in turn, change fluorescence of enzymatically generated polythymine-coated copper nanoparticles (CuNPs) (with excitation/emission maxima at 340/625 nm). The OTA-binding aptamer was immobilized on magnetic beads. When it binds OTA, it is partially released and exposes a region with a partly complimentary DNA strand (cDNA). After magnetic separation, the cDNA was employed as a primer to trigger the terminal deoxynucleotidyl transferase-mediated polymerization. This process generates polythymine which act as a template for synthesis of the CuNPs. The method is sensitive in having a 2.0 nM detection limit for OTA. It was successfully applied to the determination of OTA in spiked diluted red wine. Graphical abstract Schematic presentation of a fluorometric enhancement method for ochratoxin A assay based on ochratoxin A inducing structure switching of its aptamer and enzymatically generated polythymine for copper nanoparticles formation.