This study reported a rapid multiplex PCR assay using five primer pairs to simultaneously detect five major foodborne pathogen. All results obtained indication that the developed assay is rapid, cost-effective and reliable as higher consistency was obtained between the result of multiplex PCR and traditional culture method even there were some minor different. However, This developed method still useful for combined with traditional culture method for rapid screening of these pathogens. Therefore, the multiplex PCR assay hast a potential to be used in routine diagnostic laboratories and also mign be as a rapid screening tool in food investigation.