The invention relates to a kit for detecting food-borne pathogenic bacteria, which comprises 12 groups of primers and probes thereof, wherein in the primers, the 5' ends of downstream primers are all tagged with biotin. The kit amplifies nucleic acid in a sample to be detected by utilizing the primers, and a formed biotinylated product and a probe fixed on the surface of a chip are hybridized to form a probe-biotinylated product compound; an alkaline phosphatase-streptavidin conjugate is combined with the probe-biotinylated product compound, and then chromogenic substrate which is tetranitroblue tetrazolium chloride/5-bromine-4-chlorine-3-indolyl-phosphate (NBT/BCIP) is reacted with alkaline phosphatase to undergo a colour generation (bluish violet) reaction; and whether the sample to be detected contains the pathogenic bacteria to be detected is detected by interpreting whether a specific site has colour generation. The kit has the advantages of high detection throughput, high speed, high sensitivity, high specificity and great practicability. The invention also provides the method for detecting the food-borne pathogenic bacteria by adopting the detection kit.