Micropore Resistance Counting Platform for Multiplexed and Ultrasensitive Detection of Mycotoxins and Biomarkers

Development of a multiplexed and sensitive biosensing platform is a priority for public health security. We report a micropore resistance counting platform based on polystyrene microsphere size-based encoding and Clostridium butyricum Argonaute (CbAgo) decoding for multiplexed and ultrasensitive detection. Initially, we constructed a target DNA-modified polystyrene microsphere coding system based on micropore resistance counting. Subsequently, the precise recognition and cleavage capabilities of the guide DNA-activated CbAgo protein enable the decoding of the encoded microsphere system. Changes in the concentration of polystyrene microspheres are presented as a signal readout. The platform demonstrated excellent performance in multiplexed detection of three mycotoxins (with a sensitivity range over 4 orders of magnitude reaching the pg/mL level) and two inflammatory markers at pg/mL. Combining precise enzyme cleavage by CbAgo with micropore resistance counting, the developed platform is a multiplexed and highly sensitive detection tool with wide-ranging potential in applications such as clinical diagnosis, food safety inspection, and environmental monitoring.