Clostridium butyricum Argonaute-powered programmable platform using a magnetic nanoparticle tetrahedral DNA encoding system for the ultrasensitive and multiplexed detection of non-nucleic acid targets

Multiplexed detection of hazardous factors is important for food safety and public health. Argonaute (Ago) has been proposed as a promising multiplexed detection tool for its ability to specifically recognize and cleave target sequences. However, most reported thermophilic Ago-based sensors require high reaction temperatures and have limited applications for non-nucleic acid target detection. To overcome the temperature limitations and extend the application of Ago to multiplexed non-nucleic acid target detection, we herein report a mesophilic Clostridium butyricum Argonaute (CbAgo)-Powered Programmable (APP) platform combined with a magnetic nanoparticle tetrahedral DNA encoding system to quantitatively detect multiple non-nucleic acid targets at 37 °C. This platform achieved satisfactory results in the multiplexed detection of three mycotoxins in 30 food samples (pg/mL level) and three pathogens in 100 clinical samples (102 CFU/mL level without DNA extraction), highlighting its potential as a multiplexed and ultrasensitive detection tool in food safety, clinical diagnosis, and so forth.