Rapid detection of Salmonella in raw chicken breast using real-time PCR combined with immunomagnetic separation and whole genome amplification

Food Microbiol. 2017 May:63:111-116. doi: 10.1016/j.fm.2016.11.007. Epub 2016 Nov 21.

Abstract

We presented the first attempt to combine immunomagnetic separation (IMS), whole genome amplification by multiple displacement amplification (MDA) and real-time PCR for detecting a bacterial pathogen in a food sample. This method was effective in enabling real-time PCR detection of low levels of Salmonella enterica Serotype Enteritidis (SE) (∼10 CFU/g) in raw chicken breast without culture enrichment. In addition, it was able to detect refrigeration-stressed SE cells at lower concentrations (∼0.1 CFU/g) in raw chicken breast after a 4-h culture enrichment, shortening the detection process from days to hours and displaying no statistical difference in detection rate in comparison with a culture-based detection method. By substantially improving performance in SE detection over conventional real-time PCR, we demonstrated the potential of IMS-MDA real-time PCR as a rapid, sensitive and affordable method for detecting Salmonella in food.

Keywords: Chicken; Detection; IMS; MDA; Real-time PCR; Salmonella; Whole genome amplification.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Bacterial Load
  • DNA, Bacterial
  • Food Microbiology
  • Genome, Bacterial*
  • Immunomagnetic Separation / methods
  • Nucleic Acid Amplification Techniques
  • Poultry / microbiology*
  • Real-Time Polymerase Chain Reaction / economics
  • Real-Time Polymerase Chain Reaction / methods*
  • Salmonella enteritidis / genetics*
  • Salmonella enteritidis / growth & development
  • Salmonella enteritidis / immunology
  • Salmonella enteritidis / isolation & purification*
  • Sensitivity and Specificity

Substances

  • DNA, Bacterial