COMPARISON OF A REAL‐TIME POLYMERASE CHAIN REACTION ASSAY WITH A CULTURE METHOD FOR THE DETECTION OF SALMONELLA IN RETAIL MEAT SAMPLES

Abstract

A real‐time polymerase chain reaction (PCR) method developed in this study was compared with a conventional International Organization for Standardization culture method for the detection of Salmonella in Irish beef, chicken, pork and turkey. This also provided a snapshot of the incidence of Salmonella in such products. After selective enrichment, all meat samples (n = 100) were: (1) subjected to DNA extraction and real‐time PCR analysis using primers against Salmonella spp. specific gene (16S rRNA) or the invA virulence gene; and (2) plated on differential media and identified as Salmonella using immunological and sub‐typing methods. Retail samples (5/100) were shown to contain Salmonella using the 16S rRNA gene‐based real‐time PCR assay and the standard culture method. However, only two (of five) samples were shown to contain Salmonella using the invA gene‐based real‐time PCR assay. For the sample set examined, the developed 16S rRNA gene‐based real‐time PCR assay demonstrated comparable specificity and sensitivity to the currently used standard culture method but was considerably more rapid.

Populations and potential association of Saccharomyces cerevisiae with lactic acid bacteria in naturally fermented Korean rice wine

“…Among many available methods, real-time PCR is known to be the fastest and most reliable (30,31). Different from previous PCR methods, the protocol used herein has the merit of allowing any increasing fluorescence during gene amplification to be observed in real time through a monitor, obviating the need for subsequent electrophoresis (32), and thus free of any contamination through electrophoresis, reducing the detection time, and enabling rapid quantitative analysis.…”

Molecular beacon based real-time PCR assay for simultaneous detection of Listeria monocytogenes and Salmonella spp. in dairy products

“…over a wide range of food products including beef, sea food, fresh produce, and dairy products (Bohaychuk et al 2007;Cady et al 2005;Calvo et al 2008;Malorny et al 2007;Rodriguez-Lazaro et al 2004). However, barring a few (Bhagwat 2003;Jothikumar et al 2003), most of these assays were explored in uniplex format (Calvo et al 2008;Catarame et al 2006;De Martinis et al 2007;Krascsenicsova et al 2008;Neves et al 2008;Nguyen et al 2004;O'Grady et al 2008;Rodriguez-Lazaro et al 2004). Hence, it would be appropriate to develop multiplex assays for simultaneous detection of more than one pathogen in a food sample by targeting more than one gene specific for each of the target organisms.…”

Simultaneous detection of Listeria monocytogenes and Salmonella spp. in dairy products using real time PCR-melt curve analysis

“…in a wide range of products (Cady et al 2005;Catarame et al 2006;Bohaychuk et al 2007). However, most of these assays except a few (Bhagwat 2003;Jothikumar et al 2003) involved usage of either uniplex format or expensive probe based approaches (Nguyen et al 2004;De Martinis et al 2007;Calvo et al 2008;Krascsenicsova et al 2008;O'Grady et al 2008).…”