Development and validation of a rapid real-time PCR based method for the specific detection of Salmonella on fresh meat

Sheila McGuinness, Evonne McCabe, Edel O’Regan, Anthony Dolan, Geraldine Duffy, Catherine Burgess, Seamus Fanning, Thomas Barry, Justin O'Grady

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41 Citations (Scopus)

Abstract

In this study, a combined enrichment/real-time PCR method for the rapid detection of Salmonella on fresh meat carcasses, was designed, developed and validated in-house following requirements outlined in ISO 16140:2003. The method included an 18 h non-selective enrichment in buffered peptone water (BPW) and a 6 h selective enrichment in Rappaport Vasilliadis Soya (RVS) broth, based on the traditional culture method, ISO 6579:2002. The real-time PCR assay included an internal amplification control (IAC), was 100% specific and was sensitive to one cell equivalent. The alternative method was validated against the traditional culture method and relative accuracy of 94.9%, sensitivity of 94.7% and specificity of 100% were determined using 150 fresh meat carcass swabs. This alternative method had a detection limit of 1–10 CFU/100 cm2 for fresh meat carcass swabs and was performed in 26 h. Following further inter-laboratory studies, this alternative method could be suitable for implementation in testing laboratories for the analysis of carcass swabs.
Original languageEnglish
Pages (from-to)555-562
Number of pages8
JournalMeat Science
Volume83
Issue number3
DOIs
Publication statusPublished - 2009

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