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

Content Provider	Semantic Scholar
Author	Martinon, A. Cronin, Ultan P. Wilkinson, Martin G.
Copyright Year	2011
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.
Starting Page	485
Ending Page	496
Page Count	12
File Format	PDF HTM / HTML
DOI	10.1007/s12161-010-9188-7
Volume Number	4
Alternate Webpage(s)	http://www.bc-diagnostics.com/downloads/publications/Publikation_Martinon_EbEs_Reagent_D.pdf
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article