Integrated pumpless microfluidic chip for the detection of foodborne pathogens by polymerase chain reaction and electrochemical analysis

摘要

As food poisoning has increased interest in public hygiene, the study for a novel monitoring system for pathogen evaluation has been significantly explored. In this study, we established a simple single-chip pathogen analysis system by integrating the film-based PCR module, electrode module and the polydimethylsiloxane (PDMS)-based finger-actuated microfluidic modules. The film-based PCR module and electrode module were employed for gene amplification and electrochemical analysis, and the PDMS-based finger-actuated microfluidic modules were utilized for sample migration and mixing, which can replace external pumping systems. The integrated pumpless microfluidic chip contains the three parallel microfluidic channels for the simultaneous analysis of pathogens from multiple samples that channels are operated simultaneously by pressing the button without external pumping systems. Using the integrated pumpless microfluidic chip, the target gene of foodborne pathogens was amplified by a thermal cycling polymerase chain reaction (PCR) method in the film chamber, and the amplified gene was electrochemically quantified by using the square wave voltammetry. Escherichia coli O157∶H7 was selected as a target pathogen and the pathogen was sensitively evaluated with the limit of detection (LOD) of 10~2 colony-forming units (CFU). In regard to the significant featuring of the integrated microfluidic chip, the sensing platform developed could be used for pathogen screening as a promising tool in the field of point-of-care testing.