Validation of a 1-Day Analytical Diagnostic Real-Time PCR for the Detection of Salmonella in Different Food Meat Categor
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Validation of a 1-Day Analytical Diagnostic Real-Time PCR for the Detection of Salmonella in Different Food Meat Categories Elisabetta Delibato & Fabrizio Anniballi & Paola Sinibaldi Vallebona & Giuseppe Palleschi & Giulia Volpe & Marina Nadia Losio & Dario De Medici
Received: 31 July 2012 / Accepted: 20 January 2013 # Springer Science+Business Media New York 2013
Abstract Foodborne disease caused by Salmonella represents a worldwide public health problem. In Europe, salmonellosis is still the second most commonly recorded zoonosis. Since the standard culture method for detecting Salmonella (ISO 6579:2002) requires up to 5 days to produce results, the need to develop rapid methods represents an important issue for the authorities and the producers. The aim of the present study was the in-house validation, according to ISO 16140, of an open-formula diagnostic real-time PCR for the detection of Salmonella in all the different meat categories reported in the EU Regulations relative to microbiological criteria for food safety. The assay employed specific primers and a probe target within the ttrRSBCA locus, which allows the tetrathionate respiration in Salmonella.
E. Delibato (*) : F. Anniballi : D. De Medici Department of Food Safety and Veterinary Public Health, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy e-mail: [email protected] P. Sinibaldi Vallebona Department of Experimental Medicine and Surgery Sciences, Università “Tor Vergata”, Rome, Italy G. Palleschi : G. Volpe Department of Science and Chemical Technologies, Università “Tor Vergata”, Rome, Italy M. N. Losio Food Microbiology Unit, Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna, Brescia, Italy
Selectivity, relative accuracy, relative sensitivity and relative specificity were established by testing 110 bacterial strains and 175 various edible meat samples. Results showed 100 % selectivity, 100 % relative accuracy, 100 % relative sensitivity and 100 % relative specificity of the real-time PCR when compared to the standard culture method used as reference. In addition, in order to minimize the effect of the competitive micro-flora naturally present on meat samples, a highly nutritious and selective commercial medium (ONE Broth Salmonella, Oxoid) was evaluated in comparison with the classical non-selective pre-enrichment broth (buffered peptone water). Results demonstrated that the ONE Broth Salmonella medium increases the growth of Salmonella in the presence of competitive micro-flora. Keywords Salmonella . Real-time PCR . Validation . Meat samples
Introduction Foodborne disease caused by Salmonella represents a worldwide public health problem. In Europe, salmonellosis is still the second most commonly recorded zoonosis (EFSA 2011). Although various control and safety measures have been adopted throughout the food chain, microbiological testing of meat products, during both processing and production, still plays a significant role in preventing foodborne infections (Hoorfar 2011; Hoorfar et
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