A simple and rapid biosensor for ochratoxin A based on a structure-switching signaling aptamer

Abstract

A fast and reliable sensing platform has been developed for the detection of mycotoxin ochratoxin A (OTA) based on a target-induced structure-switching signaling aptamer. In the absence of target, a fluorescein-labeled OTA aptamer hybridizes to a complementary DNA strand containing a quencher moiety, bringing the fluorophore and the quencher into close proximity for highly efficient fluorescence quenching. Upon OTA addition, a conformational change in the aptamer releases the quencher-containing DNA strand, generating a strong concentration-dependent fluorescent signal. Using this technique, the entire detection and analysis process of OTA can be completed within 1 min. Under optimized assay conditions, a wide linear detection range (from 1 to 100 ng/mL) was achieved with a detection limit down to 0.8 ng/mL. Additionally, the proposed assay system exhibited high selectivity for OTA against other mycotoxins (aflatoxin B1 and zearalenone) and limited interference from the structural analog ochratoxin B. The biosensor was also applied to a non-contaminated corn material spiked with a dilution series of OTA, obtaining recoveries from 83% to 106%. Utilization of the proposed biosensor for quantitative determination of mycotoxins in food samples may provide significant improvements in quality control of food safety through a simple, rapid, and sensitive testing system for agricultural products monitoring.