A novel real-time polymerase chain reaction-based method for the detection and quantification of lactose-fermenting Enterobacteriaceae in the dairy and other food industries.
The presence of lactose-fermenting Enterobacteriaceae and coliforms is routinely assessed to determine the hygienic quality of water and foods, particularly dairy products. This paper reports the use of lacZ-specific primers in an SYBR green I-based real-time PCR method for the easy and rapid detection of coliforms in dairy products. A large number of bacterial species were assayed to establish the specificity of the method. The sensitivity of the method was assessed using artificially contaminated cheeses. The limit of detection was 1 coliform cell in cheese samples enriched for 8h in a culture medium. The entire procedure, including sample processing, enrichment, DNA extraction, and real-time PCR amplification, can be completed within 10 to 12h, making it a single-day assay.
Martín
M C
MC
Instituto de Productos Lácteos de Asturias, CSIC, Asturias, Spain.
Martínez
N
N
del Rio
B
B
Ladero
V
V
Fernández
M
M
Alvarez
M A
MA
D001483:Base Sequence; D003612:Dairying / Q000379:methods*; D004755:Enterobacteriaceae / Q000235:genetics / Q000302:isolation & purification*; D005285:Fermentation; D019649:Food Industry / Q000379:methods*; D007763:Lac Operon / Q000235:genetics; D007785:Lactose / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
D007785:Lactose