Multiplex PCR assay for the simultaneous detection of four common food pathogens in meat

Accepted: 11/07/2014 Abstract The study was carried out to standardize a multiplex PCR assay for the simultaneous detection of four common food pathogens such as Listeria monocytogenes, Yersinia enterocolitica, Staphylococcus aureus and Salmonella enterica Typhimurium. Pathogen-specific DNA primers targeting hly A, ail, nuc and stm genes were employed for the identification of L. monocytogenes, Y. enterocolitica, S. aureus and S. enterica Typhimurium, respectively. Specificity of primers was tested using reference strains of other bacterial strains including E. coli, E. coli O157:H7 and V. parahaemolyticus. Artificial meat inoculation was carried out to investigate the sensitivity of multiplex PCR. The results showed that the standardized multiplex PCR technique is an effective assay having high specificity for the simultaneous detection of the four target pathogens from meat. The developed technique successfully detected all four organisms inoculated together on meat at a detection sensitivity of 1cfu/g.