title: A novel method for species identification in milk and milk-based products.
abstract: This study describes a method for species-specific detection of animal DNA from different species (cattle, sheep, goat, water buffalo) in milk and dairy products. A primer set was designed in conserved region on the basis of the alignment of the sequence codifying the genomic kappa-casein gene in order to amplify all four species with a single primer pair. Polymorphisms were detected via minisequencing with extension primers designed in conserved sequences for haplotype determination that allow unambiguous assignment to each species. The method was successfully applied to the detection of raw and pasteurized milk from the four different species considered as well as to cheese products from the retail trade. Estimation of the limit of detection was carried out using a progression of dilutions of genomic DNA as well as DNA isolated from milk of a known number of somatic cells from different species in order to be able to achieve detection rates as low as 0.1% bovine milk mixed with buffalo milk.
authors.affiliation: Dipartimento S.A.V.A., Università del Molise, Via de Sanctis snc 86100 Campobasso, Italy. reale@unimol.it
authors.forename: Silvia
authors.initials: S
authors.lastname: Reale
authors.forename: Angela
authors.initials: A
authors.lastname: Campanella
authors.forename: Amalia
authors.initials: A
authors.lastname: Merigioli
authors.forename: Fabio
authors.initials: F
authors.lastname: Pilla
mesh_terms: D000818:Animals; D001483:Base Sequence; D002020:Buffaloes; D002417:Cattle; D004247:DNA / Q000302:isolation & purification*; D006041:Goats; D008892:Milk; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012756:Sheep; D013045:Species Specificity
chemical_list: D004247:DNA