title: A quadruplex PCR (qxPCR) assay for adulteration in dairy products.
abstract: This study describes the development of a quadruplex quantitative Real Time PCR (qxPCR) based on SYBR®GreenER chemistry, for rapid identification of DNA of cow, goat, sheep and buffalo in dairy products, and for quantification of cow DNA in these products. The platform was applied to: (i) mixes of milks at fixed percentages; (ii) cheeses prepared with the same mixes; (iii) commercial dairy products. The methodology enabled the detection of DNA from cow in mixes of milk and cheeses with a limit of detection (LOD) of 0.1%. When applied to commercial dairy products the qxPCR gave results comparable with each single-plex Real Time PCR. A good correlation (R(2)>0.9) between peaks' area of derivative of melting curves of amplicons and percentages of cow milk in milk mixes and cheeses, allows for an estimation of cow DNA in a dynamic range varying from 0.1-5% to 1-25%.
authors.affiliation: Department of Life Sciences, University of Parma, v.le Parco Area delle Scienze, 11/A, 43124 Parma, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Agrimonti
authors.affiliation: Department of Life Sciences, University of Parma, v.le Parco Area delle Scienze, 11/A, 43124 Parma, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Pirondini
authors.affiliation: Department of Life Sciences, University of Parma, v.le Parco Area delle Scienze, 11/A, 43124 Parma, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Marmiroli
authors.affiliation: Department of Life Sciences, University of Parma, v.le Parco Area delle Scienze, 11/A, 43124 Parma, Italy. Electronic address: nelson.marmiroli@unipr.it.
authors.forename: Nelson
authors.initials: N
authors.lastname: Marmiroli
mesh_terms: D000818:Animals; D002020:Buffaloes; D002417:Cattle; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D003611:Dairy Products / Q000032:analysis*; D005260:Female; D005504:Food Analysis / Q000379:methods*; D006041:Goats; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012756:Sheep
chemical_list: D004247:DNA
keywords: DNA quantification; Dairy products; Food frauds; Food traceability; Multiplex Real Time PCR; SYBR Green