Development of a self-priming PDMS/paper hybrid microfluidic chip using mixed-dye-loaded loop-mediated isothermal amplification assay for multiplex foodborne pathogens detection.

Foodborne pathogen is the primary cause of foodborne disease outbreak. Given its great damage, a sensitive, simple and rapid detection method is demanded. Herein, we described a self-priming polydimethylsiloxane (PDMS)/paper hybrid microfluidic chip, termed SPH chip, with mixed-dye-loaded loop-mediated isothermal amplification (LAMP) for multiplex foodborne pathogens detection. Staphylococcus aureus (SA) and Vibrio parahaemolyticus (VP) were chosen to verify the novel method. Compared to other similar detection devices, the SPH chip required easier fabrication process, less operation steps and lower cost. Additionally, the reaction result, especially for the weak-positive reaction, could be judged more accurately and conveniently due to the use of mixed-dye. Without pre-enrichment of bacteria in the food contaminated sample, the limit of detection (LOD) reached down to 1000 CFU mL-1 with high specific. Additionally, for fully exploiting the potential of SPH chip, a conceptual eight-channel detection chip was also developed. Overall, the reliable and excellent result demonstrated that the novel method had great potential to be applied in the wider range of pathogens detection or disease diagnose, especially in some resource-limited area.