title: 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.
abstract: 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.
authors.affiliation: Instituto de Productos Lácteos de Asturias, CSIC, Asturias, Spain.
authors.forename: M C
authors.initials: MC
authors.lastname: Martín
authors.forename: N
authors.initials: N
authors.lastname: Martínez
authors.forename: B
authors.initials: B
authors.lastname: del Rio
authors.forename: V
authors.initials: V
authors.lastname: Ladero
authors.forename: M
authors.initials: M
authors.lastname: Fernández
authors.forename: M A
authors.initials: MA
authors.lastname: Alvarez
mesh_terms: 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
chemical_list: D007785:Lactose