Magnetically actuated microfluidic chip combined with a G-quadruplex DNAzyme-based fluorescent/colorimetric sensor for the dual-mode detection of ochratoxin A in wheat

In this work, a portable fluorescent/colorimetric sensor based on G-quadruplex DNAzyme was constructed to achieve rapid and dual-mode detection of ochratoxin A (OTA) in wheat. OTA aptamers coupled with magnetic beads (MBs) can self-assemble with two segments of DNA and hemin to form a G-quadruplex DNAzyme structure that can catalyze the oxidation of Amplex Red (ADHP) with H2O2, making the solution red and producing strong fluorescence in solution. However, in the presence of OTA, the structure of the G-quadruplex DNAzyme was damaged, resulting in reduced catalytic activity. According to the principle of detection, a magnet-controlled chip integrating the reaction, washing, and detection was designed in this study. Shuttling the MB-DNAzyme probes onto a magnetically controlled chip considerably reduced the background signal and improved the detection efficiency and sensitivity. In addition, a portable fluorescence and colorimetric detection platform was built for on-site OTA detection.