关键词: 多重聚合酶链式反应, 食源性致病菌, 霉菌, 快速检测

Abstract: In this study, a multiplex polymerase chain reaction (PCR) method was developed to simultaneously detect Staphylococcus aureus, Listeria monocytogenes, Salmonella, Escherichia coli O157:H7, and mold in foods. Species-specific primers were designed targeting the heat-resistant nuclease gene (nuc) in S. aureus, the invader upregulation protein gene (hilA) in Salmonella, the flagellum gene (flic) in E. coli O157:H7, the virulence regulation protein gene (prfA) in L. monocytogenes and the V5 region of the 18S rRNA gene in mold, and the PCR reaction system and conditions were optimized for highly specific amplification. Sausage, bread and tofu artificially contaminated with the five food-borne pathogens were enriched at 37 ℃. The detection limits of the five food-borne pathogens within 20 hours all reached 100 CFU/25 g. The multiplex PCR method established in this study is suitable for rapid detection of common pathogenic organisms in foods. Compared with traditional detection methods, it has the advantages of rapidity, simplicity and high specificity and thus, can be used to simultaneously detect bacteria and mold in foods.

Key words: multiplex polymerase chain reaction (PCR), foodborne pathogen, mold, rapid detection