A New Validated Real-Time PCR-Based Method for the Specific and Fast Detection of Cronobacter spp. in Infant Formula |
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Authors: | Mar?al Soler Olaya Ruiz-Rueda Mireia Lopez-Siles Laia Calvó Eva Kaclíková Jesús L. García-Gil |
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Affiliation: | (1) Microbial. Sistemes i Aplicacions Anal?tiques, S.L. Parc Cient?fic i Tecnol?gic de la Universitat de Girona. Ed. Jaume Casademont, E C/Pic de Peguera, 15 E-17003 Girona, Spain;(2) Department of Microbiology and Molecular Biology, Food Research Institute, Priemyseln? 4, PO Box 25, SK-82475 Bratislava 26, Slovakia;(3) Department of Biology, Universitat de Girona, Campus de Montilivi s/n, E-17071 Girona, Spain; |
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Abstract: | In this study, a real-time polymerase chain reaction (PCR)-based method was designed for the fast detection of Cronobacter spp. (a newly proposed genus formerly known as Enterobacter sakazakii) in infant formula. The real-time PCR was positively tested with 70 Cronobacter strains, including members of all the species of this genus, and 88 non-Cronobacter strains. This new PCR-based system was validated against the reference standard ISO/TS 22964: 2006 (ISO International Organization for Standardization 2006) using 70 food matrices including powdered infant formula, follow-up formula, and hydrolyzed cereals for infants. The detection limit of the technique was found to be of 1 cfu in 10 g of food, fulfilling the requirements of the European Commission. The time of analysis, which comprises around 3–6 days using the reference method, is considerably reduced to less than 24 h using the real-time PCR-based system hereby described, allowing food industry a faster release of the stocks for commercialization. Moreover, this method includes an internal amplification control, co-amplified during each PCR run to verify the results. |
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