A quadruplex PCR (qxPCR) assay for adulteration in dairy products. (15th November 2015)

• Record Type: Journal Article
• Authors: Agrimonti, Caterina; Pirondini, Andrea; Marmiroli, Marta; Marmiroli, Nelson
• Is Part Of: Food chemistry. Volume 187(2015)
• Page Start: 58
• Page End: 64
• Publication Date: 2015-11-15
• Subjects: Multiplex Real Time PCR -- SYBR Green -- DNA quantification -- Dairy products -- Food frauds -- Food traceability; Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2015.04.017
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: A quadruplex PCR (qxPCR) assay, based on SYBRGreen was developed to test the presence of cow milk in dairy products. The qxPCR was tested on experimental milk mixes and cheeses and commercial dairy products. The capacity of qxPCR to quantify cow milk in dairy products was tested. 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%.