Abstract: To explore a fast, economical and practical multiplex PCR method which could detect Staphylococcus aureus, Salmonella, Yersinia enterocolitica common food-borne bacterial pathogens simultaneously.Three pairs of primers were designed respectively according to the nuc gene of Staphylococcus aureus, the invasive protein gene (invA) of Salmonella, the sequence of the ail gene of Yersinia enterocolitica.The specific and sensitive experiments were carried out to the amplification of single gene by PCR and Multiplex PCR amplification, and the reaction system were optimized.Three DNA fragments of 236, 475 and 127bp were amplified by three pairs of specific primers.Three food borne bacterial pathogens were simultaneously detected by the established multiplex PCR technology.The sensitivity of this method was for Staphylococcus aureus 102CFU/mL, for salmonella 102CFU/mL, for Yersinia enterocolitica 102CFU/mL.A triple PCR assay had been established for the simultaneous sample, rapid and sensitive detection of Staphylococcus aureus, salmonella, Yersinia enterocolitica in food.