title: Validation of a fast real-time PCR method to detect fraud and mislabeling in milk and dairy products.
abstract: Fast real-time PCR TaqMan assays were developed and validated for species identification in dairy products. Based on the amplification of 12S rRNA and cytB partial genes of mitochondrial DNA, the methods were demonstrated to be sensitive, fast, and species-specific for Bos taurus, Ovis aries, Bubalus bubalis, and Capra hircus. The limit of detection calculated was lower than 1%, and the efficiency was reported to be higher than 96% in every assay. An internal amplification control was used to detect possible false negatives. The method was validated by means of laboratory-prepared samples mixing different species. Moreover, 18 commercial dairy samples were analyzed by both real-time PCR and isoelectric focusing, the official European Union reference method. The 4 TaqMan assays were confirmed to be a useful tool for milk and dairy product authentication.
authors.affiliation: Ricerca e Sviluppo Biotecnologie, Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale", Via Campo Boario, 64100 Teramo, Italy. Electronic address: m.didomenico@izs.it.
authors.forename: M
authors.initials: M
authors.lastname: Di Domenico
authors.affiliation: Ricerca e Sviluppo Biotecnologie, Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale", Via Campo Boario, 64100 Teramo, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Di Giuseppe
authors.affiliation: Ricerca e Sviluppo Biotecnologie, Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale", Via Campo Boario, 64100 Teramo, Italy.
authors.forename: J D
authors.initials: JD
authors.lastname: Wicochea Rodríguez
authors.affiliation: Ricerca e Sviluppo Biotecnologie, Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale", Via Campo Boario, 64100 Teramo, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Cammà
mesh_terms: D000818:Animals; D002020:Buffaloes; D002417:Cattle; D003611:Dairy Products / Q000032:analysis*; D005504:Food Analysis / Q000379:methods*; D006041:Goats; D057230:Limit of Detection; D008892:Milk / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D034561:Sheep, Domestic; D013045:Species Specificity
keywords: dairy products; isoelectric focusing; real-time PCR; species identification