一种用于检测乳制品掺假的四重聚合酶链反应（qxPCR）检测方法。
A quadruplex PCR (qxPCR) assay for adulteration in dairy products.
作者信息
Agrimonti Caterina, Pirondini Andrea, Marmiroli Marta, Marmiroli Nelson
机构信息
Department of Life Sciences, University of Parma, v.le Parco Area delle Scienze, 11/A, 43124 Parma, Italy.
Department of Life Sciences, University of Parma, v.le Parco Area delle Scienze, 11/A, 43124 Parma, Italy.
出版信息
Food Chem. 2015 Nov 15;187:58-64. doi: 10.1016/j.foodchem.2015.04.017. Epub 2015 Apr 13.
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%.
本研究描述了一种基于SYBR®GreenER化学的四重定量实时荧光定量PCR（qxPCR）方法的开发，用于快速鉴定乳制品中牛、山羊、绵羊和水牛的DNA，并对这些产品中的牛DNA进行定量。该平台应用于：（i）固定百分比的牛奶混合物；（ii）用相同混合物制备的奶酪；（iii）商业乳制品。该方法能够检测牛奶和奶酪混合物中牛的DNA，检测限（LOD）为0.1%。当应用于商业乳制品时，qxPCR的结果与每个单重实时荧光定量PCR相当。扩增子熔解曲线导数峰面积与牛奶混合物和奶酪中牛奶百分比之间具有良好的相关性（R(2)>0.9），可以在0.1 - 5%至1 - 25%的动态范围内估计牛DNA。
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