title: Salmonella detection in poultry samples. Comparison of two commercial real-time PCR systems with culture methods for the detection of Salmonella spp. in environmental and fecal samples of poultry.
abstract: METHODS
  The efficiency of two commercial PCR methods based on real-time technology, the foodproof® Salmonella detection system and the BAX® PCR Assay Salmonella system was compared to standardized culture methods (EN ISO 6579:2002 - Annex D) for the detection of Salmonella spp. in poultry samples.
  
  
  METHODS
  Four sample matrices (feed, dust, boot swabs, feces) obtained directly from poultry flocks, as well as artificially spiked samples of the same matrices, were used. All samples were tested for Salmonella spp. using culture methods first as the gold standard. In addition samples spiked with Salmonella Enteridis were tested to evaluate the sensitivity of both PCR methods. Furthermore all methods were evaluated in an annual ring-trial of the National Salmonella Reference Laboratory of Germany.
  
  
  RESULTS
  Salmonella detection in the matrices feed, dust and boot swabs were comparable in both PCR systems whereas the results from feces differed markedly. The quality, especially the freshness, of the fecal samples had an influence on the sensitivity of the real-time PCR and the results of the culture methods. In fresh fecal samples an initial spiking level of 100cfu/25g Salmonella Enteritidis was detected. Two-days-dried fecal samples allowed the detection of 14cfu/25g. Both real- time PCR protocols appear to be suitable for the detection of Salmonella spp. in all four matrices. The foodproof® system detected eight samples more to be positive compared to the BAX® system, but had a potential false positive result in one case. In 7-days-dried samples none of the methods was able to detect Salmonella likely through letal cell damage.
  
  
  CONCLUSIONS
  In general the advantage of PCR analyses over the culture method is the reduction of working time from 4-5 days to only 2 days. However, especially for the analysis of fecal samples official validation should be conducted according to the requirement of EN ISO6579:2002 - Annex D.
authors.affiliation: Clinic for Birds, Reptiles, Amphibians and Fish, Justus-Liebig-University, Giessen, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Sommer
authors.forename: D
authors.initials: D
authors.lastname: Enderlein
authors.forename: A
authors.initials: A
authors.lastname: Antakli
authors.forename: H
authors.initials: H
authors.lastname: Schönenbrücher
authors.forename: J
authors.initials: J
authors.lastname: Slaghuis
authors.forename: T
authors.initials: T
authors.lastname: Redmann
authors.forename: M
authors.initials: M
authors.lastname: Lierz
mesh_terms: D000821:Animal Feed / Q000382:microbiology; D000818:Animals; D004391:Dust; D004783:Environmental Microbiology; D005243:Feces / Q000382:microbiology*; D011200:Poultry; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012475:Salmonella / Q000235:genetics* / Q000302:isolation & purification*
chemical_list: D004391:Dust