Real time PCR gene profiling and detection of Salmonella using a novel target: The siiA gene
Résumé
The objective of this study was to develop and evaluate a SYBR Green real time PCR method for the specific
detection of Salmonella spp using a novel target, the siiA gene. Primer specificity testing was done on a panel of
76 Salmonella strains and 32 non-Salmonella strains. The primers directed against the siiA gene amplified all
Salmonella strains tested, while non-Salmonella strains were not amplified. The melting temperatures of the
107 bp amplicons were consistently specific as they gave melting peaks around 75.5 °C. The precision of the
assay, based on intra and inter-run variations, was shown to be widely acceptable. In the second part of this
study, 45 Salmonella strains were screened for the presence of 6 virulence-associated genes (sopB, cat2, safC,
sefB and SC1248) located in several Salmonella Pathogenicity Islands (SPIs) and the spvC gene from the Salmonella
virulence plasmid. The prevalence of these genes ranged from51% to 100%. Variable virulence gene profileswere
obtained even within the same serotype.