A real-time PCR for the detection of Salmonella Enteritidis in poultry meat and consumption eggs

Multiplex TaqMan Real-Time PCR (qPCR) Assay Targeting prot6E and invA Genes for Fast and Accurate Detection of Salmonella Enteritidis

“…The qPCR developed here as opposed to an invA single target qPCR method (Feder et al, 2001;Malorny et al, 2004;Bohaychuk et al, 2007;Gonzalez-Escalona et al, 2009) is able to detect specifically SE strains by the use of an SE specific marker (prot6E). Additionally it is capable to detect other SE that might lack the prot6E gene (Malorny et al, 2007a), such as the case for SE-10. The lack of prot6E in SE strains has been co-related with the absence of the SE virulence plasmid (~ 55 kb) (Malorny et al, 2007a).…”

“…Additionally it is capable to detect other SE that might lack the prot6E gene (Malorny et al, 2007a), such as the case for SE-10. The lack of prot6E in SE strains has been co-related with the absence of the SE virulence plasmid (~ 55 kb) (Malorny et al, 2007a). Due to the importance of that plasmid in SE virulence (Bakshi et al, 2003), without it SE has a diminish virulence, and such could be the case for SE-10 which was isolated from chicken.…”

“…The product of this gene is essential for the organism's ability to invade mammalian cells and subsequently cause disease (Galan and Curtiss, III, 1991;Galan JE et al, 1992). In the case of SE in specific, several PCR and isothermal methodologies has also been developed targeting different genes (Seo et al, 2004;Malorny et al, 2007a;O'Regan et al, 2008;Hadjinicolaou et al, 2009). Although isothermal amplification techniques has some advantages over qPCR, such as increased detection limit and lower cost; still has the disadvantage that a single target can be used at a time and lacks internal control for monitoring possible inhibitors of the reaction that might exist in the food matrix analyzed.…”

Salmonella Detection Methods for Food and Food Ingredients

“…Depending on the number of targets, the analysis is carried out by a single amplification reaction of four-to five targets, or could take place via a two-step amplification reaction for five-six targets or more (Settanni and Corsetti 2007). For example, Malorny et al (2007) developed an assay for the specific detection of S. Enteritidis in whole chicken carcass rinses and consumption eggs. The assay used specifically designed primers and a TaqMan probe to target the Prot6e gene located on the S. Enteritidis 60-kb virulence plasmid.…”

Quantitative studies of the regular distribution pattern for Salmonella enteritidis in the internal organs of mice after oral challenge by a specific real-time polymerase chain reaction

“…With this assay, it is possible to carry out a rapid quantitative analysis of DNA over a wide linear range, with an unknown template. The specific DNA fragment (Sdf I) of S. enteritidis was reported by Agron et al [12] , which was screened for using the Supression Subtractive Hybridization method, and appears to only be found in serovar enteritidis strains [12,13] . Here, based on the study of Agron et al, we developed a standard curve (this methodology is the first time established which based upon the Sdf I DNA fragment), and then applied this to the study of internal organ distribution of S. enteritidis in mice.…”