Galstyan Aram G, Semipyatniy Vladislav K, Mikhailova Irina Yu, Gilmanov Khamid Kh, Bigaeva Alana V, Vafin Ramil R
All-Russian Research Institute of the Dairy Industry, Lusinovskaya str. 35, 115093 Moscow, Russia.
All-Russian Scientific Research Institute of Brewing, Beverage and Wine Industry, Rossolimo str. 7, 119021 Moscow, Russia.
Foods. 2021 Mar 11;10(3):595. doi: 10.3390/foods10030595.
DNA authentication of wines is a process of verifying their authenticity by genetic identification of the main plant component. The sample preparation of experimental and commercial wines was carried out by precipitation of wine debris by centrifugation with preliminary exposure with precipitators and co-precipitators, including developed macro- and micro-volume methods applicable to white or red wines, using polyvinylpyrrolidone as a co-precipitator. Addition of 2-mercaptoethanol and proteinase K to the lysing solution made it possible to adapt the technology for DNA extraction from the precipitated wine debris. The additionally tested technique of DNA extraction from wine debris by dimethyl sulfoxide (DMSO) lysis had fewer stages and, consequently, a lower risk of contamination. The results of further testing of one of the designed primer pairs (UFGT-F1 and UFGT-R1) in conjunction with the tested methods of wine material sample preparation and nucleic acid extraction, showed the advantage in the given set of oligonucleotides over previously used ones in terms of sensitivity, specificity and reproducibility. The developing strategy for genetic identification of grape varieties and DNA authentication of wines produced from them based on direct sequencing of polymerase chain reaction (PCR) products is implemented by interpreting the detected polymorphic positions of variable L. gene locus with distribution and split into 13 gene-associated groups.
葡萄酒的DNA鉴定是通过对主要植物成分进行基因鉴定来验证其真伪的过程。实验用葡萄酒和商业葡萄酒的样品制备是通过离心沉淀葡萄酒残渣来进行的，预先使用沉淀剂和共沉淀剂，包括适用于白葡萄酒或红葡萄酒的已开发的大体积和小体积方法，使用聚乙烯吡咯烷酮作为共沉淀剂。在裂解液中添加2-巯基乙醇和蛋白酶K使得从沉淀的葡萄酒残渣中提取DNA的技术得以适用。通过二甲基亚砜（DMSO）裂解从葡萄酒残渣中提取DNA的额外测试技术步骤更少，因此污染风险更低。对设计的一对引物（UFGT-F1和UFGT-R1）与测试的葡萄酒材料样品制备和核酸提取方法相结合进行的进一步测试结果表明，在给定的一组寡核苷酸中，其在灵敏度、特异性和可重复性方面优于先前使用的引物。基于聚合酶链反应（PCR）产物的直接测序对葡萄品种进行基因鉴定以及对用其生产的葡萄酒进行DNA鉴定的发展策略，是通过解释检测到的可变L.基因座的多态性位置并将其分布和划分为13个基因相关组来实现的。
