Portable centrifugal microfluidics system for on-site detection of foodborne pathogens.

Rapid and accurate detection of foodborne pathogens is essential to prevent outbreaks of foodborne illness and reduce public health threats. However, conventional diagnostic methods remain inadequate for on-site food safety monitoring due to low sensitivity, bulky equipment, and complex operation. Therefore, the development of portable, miniaturized, and user-friendly point-of-care testing systems has become essential. In this study, we report an all-in-one microfluidic centrifugal disc (AMCD) platform that integrates recombinase polymerase amplification (RPA) and CRISPR-Cas12a detection for rapid on-site detection of foodborne pathogens. The AMCD platform utilizes centrifugal force to automate the sequential release, reagent mixing, amplification, and detection of foodborne pathogen DNA after sample loading. Notably, the disc was configured with compartmentalized chambers to maintain physical separation of the reagents and prevent premature mixing between sequential reaction steps. To demonstrate the practical applicability of the AMCD platform, we performed multiplexed detection of four different foodborne pathogens (Staphylococcus aureus, Salmonella enteritidis, Listeria monocytogenes, and Bacillus cereus) spiked in real food samples, successfully completing the entire analysis within 60 min. Furthermore, integrating the platform with a smartphone application enables immediate on-site monitoring. The developed portable AMCD platform provides a promising approach for rapid detection of foodborne pathogens and on-site food safety monitoring.