Development and validation of a rapid real-time PCR based method for the specific detection of Salmonella on fresh meat |
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| Authors: | Sheila McGuinness Evonne McCabe Edel O’Regan Anthony Dolan Geraldine Duffy Catherine Burgess Seamus Fanning Thomas Barry Justin O’Grady |
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| Affiliation: | 1. Molecular Diagnostics Research Group, National Centre for Biomedical Engineering Science, National University of Ireland Galway, Galway, Ireland;2. Microbiology, School of Natural Sciences, National University of Ireland Galway, Galway, Ireland;3. Food Safety Department, Ashtown Food Research Centre, Teagasc, Dublin, Ireland;4. School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Dublin, Ireland |
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| 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. |
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| Keywords: | Salmonella Real-time PCR ssrA Gene IAC Fresh meat Detection |
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