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 language | English |
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Pages (from-to) | 485-496 |
Number of pages | 12 |
Journal | Food Analytical Methods |
Volume | 4 |
Issue number | 4 |
DOIs | |
Publication status | Published - Dec 2011 |
Keywords
- Detection
- Enterobacteriaceae
- Infant formula milk
- LUX™ primers
- Real-time PCR