Rapid detection of three mycotoxins in animal feed materials using competitive ELISA-based origami microfluidic paper analytical device (μPAD)

We report the development of a competitive ELISA-based origami microfluidic paper-based analytical device (μPAD) for the detection of mycotoxins in animal feed material. The μPAD was patterned using the wax printing technique with the design of a testing pad in the middle and two absorption pads at the side. Anti-mycotoxin antibodies were effectively immobilized on chitosan–glutaraldehyde-modified sample reservoirs in the μPAD. The determination of zearalenone, deoxynivalenol, and T-2 toxin in corn flour was successfully achieved by performing competitive ELISA on the μPAD in 20 min. Colorimetric results were easily distinguished by the naked eye with a detection limit of 1 µg/mL for all three mycotoxins. The μPAD integrated with competitive ELISA holds potential for practical applications in the livestock industry for rapid, sensitive, and cost-effective detection of different mycotoxins in animal feed materials. Graphical abstract