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

• Published in: Analytica chimica acta Vol. 1040; pp. 81 - 89
• Main Authors: Pang, Bo, Fu, Kaiyue, Liu, Yushen, Ding, Xiong, Hu, Jiumei, Wu, Wenshuai, Xu, Kun, Song, Xiuling, Wang, Juan, Mu, Ying, Zhao, Chao, Li, Juan
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 21.12.2018; Elsevier BV
• Subjects: Bacteria; Bioassays; Coloring Agents - chemistry; Dimethylpolysiloxanes - chemistry; Disease control; Dyes; Fabrication; Food contamination; Food safety; Foodborne diseases; Foodborne pathogen; Foodborne pathogens; Gene amplification; Loop-mediated isothermal amplification; Microfluidic Analytical Techniques; Microfluidic chip; Microfluidics; Mixed dye; Multiplex detection; Multiplexing; Outbreaks; Pathogens; Polydimethylsiloxane; Priming; Semiconductors; Silicone resins; Staphylococcus aureus - isolation & purification; Staphylococcus aureus - pathogenicity; Temperature; Vibrio parahaemolyticus - isolation & purification; Vibrio parahaemolyticus - pathogenicity; Waterborne diseases; Microfluidic chip; Loop-mediated isothermal amplification; Food safety; Mixed dye; Multiplex detection; Foodborne pathogen
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2018.07.024

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. [Display omitted] •A low-cost SPE chip combining paper and PDMS was developed.•Multiplex foodborne pathogens could be simultaneously detected using SPE chip.•Self-priming introduction of SPE chip made the operation simpler.•Mixed-dye-loaded LAMP assay made the result more accurate and distinguishable.•The practicability of this method has been verified using spiked samples.