Development of a multiplex PCR system for the simultaneous detection of bacillus cereus, salmonella spp. and type A enterotoxigenic staphylococcus aureus in food samples

• Main Authors: King, Jen-Yun, 金仁筠
• Other Authors: Hau-Yang Tsen
• Format: Others
• Language: zh-TW
• Published: 1996
• Online Access: http://ndltd.ncl.edu.tw/handle/82877532801689884511

碩士 === 國立中興大學 === 食品科學系 === 84 === Bacillus cereus,Salmonella spp. and type A enterotoxigenic Staphylococcus aureus are common food-bornepathogenic bacteria. Conventional microbiologicalmethods for the detection of these bacteria normally take 5-7 days.Therefore,rapid methods for the detection of these bacteriaare important. In this study, we at first developed PCR todetect Salmonella spp. in feed and the intestinal content of chicken. Prior to the PCR enrichmenet step was used. The PCR primers TSII / TS5 would allow the amplification of a 375bp PCR product. The deection sensitivity was 10 CFU/ml. Although the PCR detection systems for Bacillus cereus,Salmonella spp. and type A enterotoxigenic Staphylococcus aureus have been reported. PCR system for the simultaneousdetection of these bacteria has not been reported, Since PCR method is one of the rapidest methods for the detection offood pathogens. In this report, We try to develope a multiplex PCR system for the simultaneous detection of B.cereus, Salmonella and type A enterotoxigenic S. aureus infood samples. Prior to the PCR assay, the targer cells mustbe enriched,For exanples type A enterotoxigenic Staphylococcus aureus, Salmonella and B. cereus could beenriched using media of 10% NaCl TSB, CTET and TSPB,repectively. The PCR conditions used are 94 J 20sec, 62 J 25sec, 72 J 30sec, with 40 PCR cycles. For this multipex PCRsystem, the primers used were A1 /A2, TSII / TS5 and Pf /Cr, and the primers ratio are 40 pmol: 3 pmol: 50 pmol. When target bacteria in cooked rice we are detected by method described above, the detection sensitivity was 10 CFU / g.