Nuclease fluorescence assay for the detection of verotoxin genes in raw milk.
OBJECTIVE
To develop a rapid, high throughput PCR method for the detection of verotoxigenic Escherichia coli (VTEC) in raw milk based on TaqMan PCR.


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.


CONCLUSIONS
The TaqMan PCR offers a rapid semiautomated alternative to conventional PCR methods for the detection of VTEC in raw milk.
Bürk
Christine
C
Institute for Hygiene and Technology of Food of Animal Origin, University of Munich, Germany. C.Buerk@mh.vetmed.uni-muenchen.de
Braumiller
I G B
IG
Becker
H
H
Märtlbauer
E
E
D000818:Animals; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D003851:Deoxyribonucleases / Q000032:analysis*; D005453:Fluorescence; D005798:Genes, Bacterial; D008892:Milk / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D022603:Shiga Toxins / Q000032:analysis / Q000235:genetics*; D013045:Species Specificity; D019914:Taq Polymerase / Q000378:metabolism*
D004269:DNA, Bacterial; D022603:Shiga Toxins; D019914:Taq Polymerase; D003851:Deoxyribonucleases