The development of rapid, infallible, and sensitive methods of detecting foodborne pathogens has received much impetus in recent years owing to an increased public awareness of the health hazards. For the rapid and simultaneous detection of these foodborne pathogens, a multiplex PCR method was developed. Yersinia enterocolitica, Staphylococcus aureus, and Shigella spp. are bacteria of concern because of their specific growing condition that enables them to live at low temperatures. In order to detect each pathogenic bacterium, specific primers from Y. enterocolitica, St. aureus, and Sh. flexneri were selected and validated successfully. To apply this method to food stored at low temperature, Y. enterocolitica, St. aureus, and Sh. flexneri were artificially inoculated in lettuce and incubated for enrichment. The multiplex PCR assays were able to simultaneously detect three pathogens, and the presence of three bands was observed at initial inoculation levels of approximately 1$ imes$10$^1$ CFU/g in lettuce. Therefore, this method could be used for simultaneous detection of Y. enterocolitica, St. aureus, and Shigella spp. contaminated in lettuce during cultivation, transportation, preservation, and storage.