Demonstration of the effective performance of a combined enrichment/real-time PCR method targeting the prfA gene of Listeria monocytogenes by testing fresh naturally contaminated acid curd cheese

Lett Appl Microbiol. 2010 Oct;51(4):480-4. doi: 10.1111/j.1472-765x.2010.02925.x.

Abstract

Aims: A rapid real-time PCR-based method for the detection of Listeria monocytogenes was applied to the examination of 44 Quargel cheese samples from a recent outbreak in Austria and compared to the standard method according to ISO-16140.

Methods and results: The combined enrichment/real-time PCR method amplifying the prfA locus was performed according to [Rossmanith et al.(2006) Res Microbiol, 157, 763-771]. Qualitative and quantitative examination of the samples was performed according to the standard method ISO-11290. Comparison of the combined enrichment/real-time PCR method with ISO-11290 resulted in 100% relative accuracy, 100% relative sensitivity and 100% relative specificity.

Conclusions: A previously published study describing the validation of the method, including samples after storage at -80 degrees C, resulted in lower performance values. In contrast, the samples were stored at +4 degrees C in this study. The results of this study indicate an effect of storage, thus masking the true performance of the method.

Significance and impact of the study: The results of this study are discussed together with the previously published data to demonstrate the excellent qualities of this rapid (< or = 30 h) method when applied to fresh specimens stored at +4 degrees C.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Austria
  • Bacterial Proteins / genetics*
  • Bacteriological Techniques
  • Cheese / microbiology*
  • Colony Count, Microbial / methods
  • Culture Media
  • DNA, Bacterial / analysis
  • DNA, Bacterial / genetics
  • Food Contamination / analysis*
  • Food Microbiology
  • Humans
  • Listeria monocytogenes / genetics
  • Listeria monocytogenes / growth & development
  • Listeria monocytogenes / isolation & purification*
  • Peptide Termination Factors / genetics*
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Time Factors

Substances

  • Bacterial Proteins
  • Culture Media
  • DNA, Bacterial
  • Peptide Termination Factors
  • PrfA protein, Listeria monocytogenes