AIMS: To develop a rapid, high throughput PCR method for the detection of verotoxigenic Escherichia coli (VTEC) in raw milk based on TaqMan PCR. METHODS AND RESULTS: Two TaqMan PCR systems for the detection of verotoxin genes 1 and 2, respectively, have been established. A total of 74 bacterial strains, among them 15 VTEC, were used to characterize the PCR tests. No false negative and no false positive reactions were observed. When artificially contaminated raw milk samples of 25 ml were cultured in enrichment broth for 24 h, inocula of 10(-1) cells ml-1 could be detected. CONCLUSIONS: The TaqMan PCR systems are feasible for the detection of VTEC in raw milk. SIGNIFICANCE AND IMPACT OF THE STUDY: The TaqMan PCR offers a rapid semiautomated alternative to conventional PCR methods for the detection of VTEC in raw milk.
AIMS: To develop a rapid, high throughput PCR method for the detection of verotoxigenic Escherichia coli (VTEC) in raw milk based on TaqMan PCR. METHODS AND RESULTS: Two TaqMan PCR systems for the detection of verotoxin genes 1 and 2, respectively, have been established. A total of 74 bacterial strains, among them 15 VTEC, were used to characterize the PCR tests. No false negative and no false positive reactions were observed. When artificially contaminated raw milk samples of 25 ml were cultured in enrichment broth for 24 h, inocula of 10(-1) cells ml-1 could be detected. CONCLUSIONS: The TaqMan PCR systems are feasible for the detection of VTEC in raw milk. SIGNIFICANCE AND IMPACT OF THE STUDY: The TaqMan PCR offers a rapid semiautomated alternative to conventional PCR methods for the detection of VTEC in raw milk.