Rapid Detection of Foodborne Pathogenic Microorganisms Based on Recombinase Polymerase Amplification and Lateral Chromatography Microfluidic Chip Technology

ABSTRACT Background Developing highly sensitive, simple, and rapid detection techniques for the accurate detection of foodborne pathogens is critical for food safety. This study designed a microfluidic chip integrating recombinase polymerase amplification (RPA) and colloidal gold lateral chromatography for the visual detection of three high‐risk foodborne pathogens: Escherichia coli O157:H7, Vibrio parahaemolyticus (VP), and Vibrio cholerae (VC). Methods A lateral flow microfluidic chip was fabricated by integrating a chromatography test strip with a laser‐ablated microfluidic structure. RPA primers targeting the rfbE gene of E. coli O157:H7, the toxR gene of VP, and the ctxA gene of V. cholerae were designed. The RPA products were validated by agarose gel electrophoresis, followed by optimization of the primer concentration, temperature, and reaction time. Results The chip detected pathogens via RPA (at 40°C for 20 min) and a visual readout was obtained within 10 min, achieving 10 2 CFU/mL detection sensitivity for all three pathogens with high specificity, reproducibility, and stability. Conclusion This microfluidic chip‐based method enables portable on‐site foodborne pathogen detection without the need for costly thermal cyclers/signal readers, thereby maintaining accuracy and affordability.