Comparison of In-house and Commercial Real-time PCR Systems for the Detection of Enterobacteriaceae and their Evaluation Within an Interlaboratory Study Using Infant Formula Samples

Alice Martinon, Ultan P. Cronin, Martin G. Wilkinson

Research output: Contribution to journalArticlepeer-review

Abstract

Traditional detection methods for Enterobacteriaceae in foods are time-consuming and laborious. The current study assessed the specificity of three real-time PCR primer sets. Set A (IEC primers) targeted the conserved flanking regions of the 16S rRNA, the 16S-ITS-23S gene region. Set B (ENT primers) annealed to Escherichia coli 16S ribosomal RNA gene. The third set (C) used a D-LUX™ (Light Upon eXtension) single FAM-labelled forward primer and a corresponding unlabeled primer. Set A was specific for E. coli and for some non-Enterobacteriaceae. SYBR Green-based real-time PCR confirmed the specificity of set B for the Enterobacteriaceae but also detected Vibrionaceae. In contrast, set C was poorly specific. However, set D including the forward LUX™ primer from set C and the reverse primer from set B had a specificity comparable to that of set B, but with higher sensitivity. This combined set was successfully applied to detect Enterobacteriaceae in infant milk formula and compared favourably with a commercial real-time PCR kit.

Original languageEnglish
Pages (from-to)485-496
Number of pages12
JournalFood Analytical Methods
Volume4
Issue number4
DOIs
Publication statusPublished - Dec 2011

Keywords

  • Detection
  • Enterobacteriaceae
  • Infant formula milk
  • LUX™ primers
  • Real-time PCR

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