Pereira Leonor, Gomes Sónia, Castro Cláudia, Eiras-Dias José Eduardo, Brazão João, Graça António, Fernandes José R, Martins-Lopes Paula
University of Trás-os-Montes and Alto Douro, P.O. Box 1013, 5000-911 Vila Real, Portugal; University of Lisboa, Faculty of Sciences, BioISI - Biosystems & Integrative Sciences Institute, Campo Grande, 1749-016 Lisboa, Portugal.
National Institute for Agricultural and Veterinary Research (INIAV), 2565-191 Dois Portos, Portugal.
Food Chem. 2017 Feb 1;216:80-6. doi: 10.1016/j.foodchem.2016.07.185. Epub 2016 Aug 1.
Wine authenticity methods are in increasing demand mainly in Denomination of Origin designations. The DNA-based methodologies are a reliable means of tracking food/wine varietal composition. The main aim of this work was the study of High Resolution Melting (HRM) application as a screening method for must and wine authenticity. Three sample types (leaf, must and wine) were used to validate the three developed HRM assays (Vv1-705bp; Vv2-375bp; and Vv3-119bp). The Vv1 HRM assay was only successful when applied to leaf and must samples. The Vv2 HRM assay successfully amplified all sample types, allowing genotype discrimination based on melting temperature values. The smallest amplicon, Vv3, produced a coincident melting curve shape in all sample types (leaf and wine) with corresponding genotypes. This study presents sensitive, rapid and efficient HRM assays applied for the first time to wine samples suitable for wine authenticity purposes.
葡萄酒真伪鉴定方法的需求主要集中在原产地命名产区。基于DNA的方法是追踪食品/葡萄酒品种组成的可靠手段。这项工作的主要目的是研究高分辨率熔解曲线分析（HRM）作为葡萄汁和葡萄酒真伪筛选方法的应用。使用了三种样品类型（叶片、葡萄汁和葡萄酒）来验证所开发的三种HRM分析方法（Vv1-705bp；Vv2-375bp；和Vv3-119bp）。Vv1 HRM分析方法仅在应用于叶片和葡萄汁样品时成功。Vv2 HRM分析方法成功扩增了所有样品类型，能够根据熔解温度值进行基因型区分。最小的扩增子Vv3在所有样品类型（叶片和葡萄酒）中产生了与相应基因型一致的熔解曲线形状。本研究首次展示了适用于葡萄酒真伪鉴定目的的葡萄酒样品的灵敏、快速且高效的HRM分析方法。
