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%.

A quadruplex PCR (qxPCR) assay for adulteration in dairy products / Agrimonti, Caterina; Pirondini, Andrea; Marmiroli, Marta; Marmiroli, Nelson. - In: FOOD CHEMISTRY. - ISSN 0308-8146. - 187(2015), pp. 58-64. [10.1016/j.foodchem.2015.04.017]

A quadruplex PCR (qxPCR) assay for adulteration in dairy products

AGRIMONTI, Caterina;PIRONDINI, Andrea;MARMIROLI, Marta;MARMIROLI, Nelson
2015

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%.
A quadruplex PCR (qxPCR) assay for adulteration in dairy products / Agrimonti, Caterina; Pirondini, Andrea; Marmiroli, Marta; Marmiroli, Nelson. - In: FOOD CHEMISTRY. - ISSN 0308-8146. - 187(2015), pp. 58-64. [10.1016/j.foodchem.2015.04.017]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11381/2795154
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