Detection of Listeria monocytogenes using a commercial PCR kit and different DNA extraction methods |
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| Affiliation: | 1. Environment and Biotechnology Centre, Faculty of Life and Social Sciences, Swinburne University of Technology, PO Box 218, Hawthorn 3122 Victoria, Australia;2. bioMérieux Australia Pty Ltd, Unit 25 Parkview Business Centre, 1 Maitland Place, Baulkham Hills, NSW 2153, Australia;1. Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità, Viale Regina Elena, 299, 00161 Rome, Italy;2. Instituto Tecnológico Agrario de Castilla y León (ITACyL), Valladolid, Spain;3. Microbiology Section, Faculty of Sciences, University of Burgos, Burgos, Spain;4. Istituto Zooprofilattico Sperimentale delle Venezie, SCS 8 — Valorizzazione delle Produzioni Alimentari, Padua, Italy;5. Alma Mater Studiorum, Università di Bologna, Dip. Scienze e Tecnologie Agro-Alimentari Ozzano dell''Emilia (BO), Italy;7. National Food Chain Safety Office, Food and Feed Safety Directorate, Food Microbiological National Reference Laboratory, Mester, Budapest, Hungary;8. CNTA, Centro Nacional de Tecnología y Seguridad Alimentaria, San Adrian, Navarra, Spain;9. University of Zagreb, Faculty of Veterinary Medicine, Dpt. of Hygiene, Technology and Food Safety, Zagreb, Croatia;10. Section for Bacteriology — Food and GMO, Norwegian Veterinary Institute, Oslo, Norway;11. Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark;12. Direzione Operativa Controllo degli Alimenti, Istituto Zooprofilattico Sperimentale del Lazio e Toscana, Rome, Italy;13. Istituto Zooprofilattico Sperimentale della Lombardia e dell''Emilia Romagna, Brescia, Italy |
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| Abstract: | The aim of our work was to evaluate a new commercial test kit for the detection of Listeria monocytogenes by PCR, using different DNA extraction methods. Food samples (pork sausage and “mozzarella” cheese) were spiked with known concentrations of L. monocytogenes and culture-enriched for 24 h. DNA extracted using three commercial kits and two standard methods, was amplified in species-specific PCR employing a L. monocytogenes PCR Detection Kit (Diatheva). The PCR-based method proved to be a reliable means of detecting the pathogen in food samples independently from the extraction procedure used, even for a contamination cell number of 1 cfu/g before culture enrichment. The molecular assay, showing perfect agreement with standard microbiological tests and a considerably shortened analysis time, provides a sensitive and rapid alternative for applications in the testing of foods for microbiological contamination, and highlights the potential of PCR technology in routine food control. |
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