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A novel method for species identification in milk and milk-based products

Published online by Cambridge University Press:  29 January 2008

Silvia Reale*
Affiliation:
Dipartimento S.A.V.A., Università del Molise, Via de Sanctis snc86100 Campobasso, Italy Parco Scientifico e Tecnologico del Molise “Moliseinnovazione” S.C.a r.l., Via de Sanctis snc86100 Campobasso, Italy
Angela Campanella
Affiliation:
Dipartimento S.A.V.A., Università del Molise, Via de Sanctis snc86100 Campobasso, Italy
Amalia Merigioli
Affiliation:
Parco Scientifico e Tecnologico del Molise “Moliseinnovazione” S.C.a r.l., Via de Sanctis snc86100 Campobasso, Italy
Fabio Pilla
Affiliation:
Dipartimento S.A.V.A., Università del Molise, Via de Sanctis snc86100 Campobasso, Italy
*
*For correspondence; e-mail: [email protected]

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 κ-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.

Type
Research Article
Copyright
Copyright © Proprietors of Journal of Dairy Research 2008

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