title: European validation of Real-Time PCR method for detection of Salmonella spp. in pork meat.
abstract: The classical microbiological method for detection of Salmonella spp. requires more than five days for final confirmation, and consequently there is a need for an alternative methodology for detection of this pathogen particularly in those food categories with a short shelf-life. This study presents an international (at European level) ISO 16140-based validation study of a non-proprietary Real-Time PCR-based method that can generate final results the day following sample analysis. It is based on an ISO compatible enrichment coupled to an easy and inexpensive DNA extraction and a consolidated Real-Time PCR assay. Thirteen laboratories from seven European Countries participated to this trial, and pork meat was selected as food model. The limit of detection observed was down to 10 CFU per 25 g of sample, showing excellent concordance and accordance values between samples and laboratories (100%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (100%) when the results obtained for the Real-Time PCR-based methods were compared to those of the ISO 6579:2002 standard method. The results of this international trial demonstrate that the evaluated Real-Time PCR-based method represents an excellent alternative to the ISO standard. In fact, it shows an equal and solid performance as well as it reduces dramatically the extent of the analytical process, and can be easily implemented routinely by the Competent Authorities and Food Industry laboratories.
authors: Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità Viale Regina Elena, 299 00161 Rome, Italy. Electronic address: elisabetta.delibato@iss.it.
authors: Elisabetta
authors: E
authors: Delibato
authors: Instituto Tecnológico Agrario de Castilla y León (ITACyL), Valladolid, Spain; Microbiology Section, Faculty of Sciences, University of Burgos, Burgos, Spain.
authors: David
authors: D
authors: Rodriguez-Lazaro
authors: Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità Viale Regina Elena, 299 00161 Rome, Italy.
authors: Monica
authors: M
authors: Gianfranceschi
authors: Alma Mater Studiorum - Università di Bologna, Dip. Scienze e Tecnologie Agro-Alimentari, Ozzano dell'Emilia, BO, Italy.
authors: Alessandra
authors: A
authors: De Cesare
authors: Istituto Zooprofilattico Sperimentale delle Venezie, Dept. Food Safety, Legnaro (PD), Italy.
authors: Damiano
authors: D
authors: Comin
authors: Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità Viale Regina Elena, 299 00161 Rome, Italy.
authors: Antonietta
authors: A
authors: Gattuso
authors: Instituto Tecnológico Agrario de Castilla y León (ITACyL), Valladolid, Spain.
authors: Marta
authors: M
authors: Hernandez
authors: Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità Viale Regina Elena, 299 00161 Rome, Italy.
authors: Michele
authors: M
authors: Sonnessa
authors: Alma Mater Studiorum - Università di Bologna, Dip. Scienze e Tecnologie Agro-Alimentari, Ozzano dell'Emilia, BO, Italy.
authors: Frédérique
authors: F
authors: Pasquali
authors: National Food Chain Safety Office, Food and Feed Safety Directorate, Food Microbiological National Reference Laboratory, Mester, Budapest, Hungary.
authors: Zuzsanna
authors: Z
authors: Sreter-Lancz
authors: CNTA, Centro Nacional de Tecnología y Seguridad Alimentaria, San Adrian, Navarra, Spain.
authors: María-José
authors: MJ
authors: Saiz-Abajo
authors: CNTA, Centro Nacional de Tecnología y Seguridad Alimentaria, San Adrian, Navarra, Spain.
authors: Javier
authors: J
authors: Pérez-De-Juan
authors: CNTA, Centro Nacional de Tecnología y Seguridad Alimentaria, San Adrian, Navarra, Spain.
authors: Javier
authors: J
authors: Butrón
authors: University of Zagreb, Faculty of Veterinary Medicine, Dept. of Poultry Diseases with Clinic, Zagreb, Croatia.
authors: Estella
authors: E
authors: Prukner-Radovcic
authors: University of Zagreb, Faculty of Veterinary Medicine, Dept. of Poultry Diseases with Clinic, Zagreb, Croatia.
authors: Danijela
authors: D
authors: Horvatek Tomic
authors: Section for Bacteriology - Food and GMO Norwegian Veterinary Institute, Oslo, Norway.
authors: Gro S
authors: GS
authors: Johannessen
authors: Department of Veterinary Disease Biology, University of Copenhagen, Denmark.
authors: Džiuginta
authors: D
authors: Jakočiūnė
authors: Department of Veterinary Disease Biology, University of Copenhagen, Denmark.
authors: John E
authors: JE
authors: Olsen
authors: Anses, Laboratoire de Ploufragan-Plouzané, Unité Hygiène et Qualité des Produits Avicoles et Porcins, Ploufragan, France.
authors: Marianne
authors: M
authors: Chemaly
authors: Anses, Laboratoire de Ploufragan-Plouzané, Unité Hygiène et Qualité des Produits Avicoles et Porcins, Ploufragan, France.
authors: Francoise
authors: F
authors: Le Gall
authors: Instituto Tecnológico Agrario de Castilla y León (ITACyL), Valladolid, Spain.
authors: Patricia
authors: P
authors: González-García
authors: Istituto Zooprofilattico Sperimentale delle Venezie, Dept. Food Safety, Legnaro (PD), Italy.
authors: Antonia Anna
authors: AA
authors: Lettini
authors: University of Zagreb, Faculty of Veterinary Medicine, Dept. of Poultry Diseases with Clinic, Zagreb, Croatia.
authors: Maja
authors: M
authors: Lukac
authors: Anses, Laboratoire de Ploufragan-Plouzané, Unité Hygiène et Qualité des Produits Avicoles et Porcins, Ploufragan, France.
authors: Segolénè
authors: S
authors: Quesne
authors: Istituto Zooprofilattico Sperimentale delle Venezie, Dept. Food Safety, Legnaro (PD), Italy.
authors: Claudia
authors: C
authors: Zampieron
authors: Direzione Operativa Controllo degli Alimenti, Istituto Zooprofilattico Sperimentale del Lazio e Toscana, Rome, Italy.
authors: Paola
authors: P
authors: De Santis
authors: Direzione Operativa Controllo degli Alimenti, Istituto Zooprofilattico Sperimentale del Lazio e Toscana, Rome, Italy.
authors: Sarah
authors: S
authors: Lovari
authors: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Brescia, Italy.
authors: Barbara
authors: B
authors: Bertasi
authors: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Brescia, Italy.
authors: Enrico
authors: E
authors: Pavoni
authors: Istituto Zooprofilattico del Sperimentale Mezzogiorno Portici, NA, Italy.
authors: Yolande T R
authors: YT
authors: Proroga
authors: Istituto Zooprofilattico del Sperimentale Mezzogiorno Portici, NA, Italy.
authors: Federico
authors: F
authors: Capuano
authors: Alma Mater Studiorum - Università di Bologna, Dip. Scienze e Tecnologie Agro-Alimentari, Ozzano dell'Emilia, BO, Italy.
authors: Gerardo
authors: G
authors: Manfreda
authors: Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità Viale Regina Elena, 299 00161 Rome, Italy.
authors: Dario
authors: D
authors: De Medici
mesh_terms: D000818:Animals; D004269:DNA, Bacterial / Q000032:analysis; D005060:Europe; D005516:Food Microbiology / Q000379:methods*; D008460:Meat / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012475:Salmonella / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013552:Swine
chemical_list: D004269:DNA, Bacterial
keywords: Artificial contamination; ISO 6579; Pork meat; Real Time PCR; Salmonella; Validation