Silver nanocluster-based aptasensor for the label-free and enzyme-free detection of ochratoxin A

Mycotoxin contamination in cereal is a global concern, threatening food safety and human health, necessitating the development of rapid on-site methods. Here, a label- and enzyme-free biosensor was developed based on aptamer-regulated DNA-silver nanoclusters (AgNCs) for rapid detection of ochratoxin A (OTA). A novel DNA-templated AgNCs emitting strong red fluorescence was designed and synthesized in this study. The partial sequence of the DNA template was selected from the complementary OTA aptamer (Apt-OTA) sequence, which can quench fluorescence from the AgNCs via hybridization in the absence of OTA. In the presence of OTA, the high OTA-Aptamer affinity prevented the Apt-OTA from quenching the AgNCs, resulting in "turn on" of the fluorescence. This biosensor eliminated the use of costly reagents, complex pretreatments, and sophisticated equipment, which could realize the point-of-care testing (POCT) of OTA with a limit of detection (LOD) of 1.3 nM and a detection time of 45 min.