title: Real time PCR gene profiling and detection of Salmonella using a novel target: The siiA gene.
abstract: 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 from 51% to 100%. Variable virulence gene profiles were obtained even within the same serotype.
authors.affiliation: Unité de recherche Toxicologie-Microbiologie Environnementale et Santé (UR11ES70), Faculté des Sciences de Sfax, Route de la Soukra km 3.5, B.P. n° 1171-3000 Sfax, Université de Sfax, Sfax, Tunisie.
authors.forename: Amal
authors.initials: A
authors.lastname: Ben Hassena
authors.affiliation: Unité de recherche Toxicologie-Microbiologie Environnementale et Santé (UR11ES70), Faculté des Sciences de Sfax, Route de la Soukra km 3.5, B.P. n° 1171-3000 Sfax, Université de Sfax, Sfax, Tunisie.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Barkallah
authors.affiliation: Unité de recherche Toxicologie-Microbiologie Environnementale et Santé (UR11ES70), Faculté des Sciences de Sfax, Route de la Soukra km 3.5, B.P. n° 1171-3000 Sfax, Université de Sfax, Sfax, Tunisie.
authors.forename: Imen
authors.initials: I
authors.lastname: Fendri
authors.affiliation: Laboratoire de Microbiologie, Département agroalimentaire Agrocampus Ouest, Rennes, France.
authors.forename: Noel
authors.initials: N
authors.lastname: Grosset
authors.affiliation: Centre Régional des Recherches Vétérinaire, Sfax, Tunisie.
authors.forename: Idriss
authors.initials: I
authors.lastname: Ben Neila
authors.affiliation: Laboratoire de Microbiologie, Département agroalimentaire Agrocampus Ouest, Rennes, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Gautier
authors.affiliation: Unité de recherche Toxicologie-Microbiologie Environnementale et Santé (UR11ES70), Faculté des Sciences de Sfax, Route de la Soukra km 3.5, B.P. n° 1171-3000 Sfax, Université de Sfax, Sfax, Tunisie.
authors.forename: Radhouane
authors.initials: R
authors.lastname: Gdoura
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D012475:Salmonella / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D012480:Salmonella Infections / Q000175:diagnosis / Q000382:microbiology; D012481:Salmonella Infections, Animal / Q000175:diagnosis / Q000382:microbiology; D044366:Transition Temperature
chemical_list: D017931:DNA Primers
keywords: Real time PCR; SPI; Salmonella detection; Virulence; spvC