title: A novel real-time PCR-based method for the detection of Listeria monocytogenes in food.
abstract: OBJECTIVE
  A new real-time PCR-based method was developed for the detection of Listeria monocytogenes in food.
  
  
  RESULTS
  A two-step enrichment involving a 24-h incubation in half-Fraser broth followed by a 6-h subculture in Fraser broth was used, followed by cell lysis and real-time PCR with primers and a TaqMan probe previously developed in our laboratory. When the method was evaluated with 144 naturally contaminated food samples, 44 were detected as positive by the PCR-based method and 42 by the standard method EN ISO 11290-1. With 61 food samples artificially contaminated at a level of 10(0) CFU per 25 g, 61 and 58 positive samples were detected by the respective methods.
  
  
  CONCLUSIONS
  The developed real-time PCR-based method facilitated the detection of L. monocytogenes in food on the next day after the sample reception, with a reduction of false-positive results because of dead bacterial cells and false-negative results because of PCR inhibitors.
  
  
  CONCLUSIONS
  The method can be used for L. monocytogenes detection in food as a faster alternative to current methods.
authors.affiliation: Department of Microbiology and Molecular Biology, Food Research Institute, Bratislava, Slovakia.
authors.forename: K
authors.initials: K
authors.lastname: Oravcová
authors.forename: T
authors.initials: T
authors.lastname: Kuchta
authors.forename: E
authors.initials: E
authors.lastname: Kaclíková
mesh_terms: D005506:Food Contamination; D005516:Food Microbiology; D008089:Listeria monocytogenes / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity