High-flux simultaneous screening of common foodborne pathogens and their virulent factors.

Abstract

Rapid and sensitive detection techniques for foodborne pathogens are important to the food industry. However, traditional detection methods rely on bacterial culture in combination with biochemical tests, a process that typically takes 4-7days to complete. In this study, we described a high-flux polymerase chain reaction (PCR) method for simultaneous detection of nine targeted genes (rfbE, stx1, stx2, invA, oprI, tlh, trh, tdh, and hlyA) with multiplex strains. The designed primers were highly specific for their respective target gene fragments. As the selected primers follow the principles of similar melting and annealing temperature, all the targeted genes could be detected for one strain with the same PCR program. Combining with 96-well PCR plate, by adding a single different gene to each well in each row, both the ATCC strains (E. coli, Salmonella spp., V. parahaemolyticus, L. monocytogenes, P. aeruginosa, S. aureus) and the clinical strains (E. coli, P. aeruginosa, S. aureus) were simultaneously detected to carry their specific and virulence genes. Therefore, using 96-well PCR plate for PCR amplification might be applied to high-flux sequencing of specific and virulence genes.