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Detection and discrimination of cereal and leguminous species in chestnut flour by duplex PCR

机译:双重PCR检测和鉴别板栗粉中的谷物和豆科物种

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Chestnut and chestnut-derived products are quite expensive and thus a possible target for fraudulent labeling. To obtain a French Label of Origin, chestnut producers need to certify the purity of their products. Chestnut-derived products are consumed by people who may suffer from celiac disease or are allergic to cereals. For these reasons, we developed a qualitative PCR-based method to detect cereal and leguminous species in chestnut flour. The presence of common wheat and barley was determined by amplifying part of the puroindoline-a gene, the presence of rye by amplifying part of the secaloindoline-a gene, the presence of durum wheat, rice, maize and chickpea by amplifying part of lipid transfer protein genes, the presence of oat by amplifying part of a thionin gene, the presence of kidney bean by amplifying part of a late embryogenesis-abundant protein, the presence of soybean by amplifying part of a lectin gene and the presence of fava bean by amplifying part of a nodulin gene. A chestnut thaumatine gene was used as control. PCR conditions were optimized to detect 1% of adulteration. Duplex PCR and specially designed sets of primers that allow amplification of several related species limited the number and cost of analyses required. Interpretation of the results is facilitated by using a decision tree.
机译:栗子和栗子衍生的产品非常昂贵,因此可能成为欺诈性标签的目标。为了获得法国原产地标记,栗子生产商需要证明其产品的纯度。可能患有乳糜泻或对谷物过敏的人食用栗子衍生产品。由于这些原因,我们开发了一种基于PCR的定性方法来检测栗粉中的谷物和豆类。通过扩增部分puroindoline-a基因来确定普通小麦和大麦的存在,通过扩增部分secaloindoline-a基因来确定黑麦的存在,通过增加部分脂质转移来确定硬粒小麦,水稻,玉米和鹰嘴豆的存在。蛋白质基因,通过扩增部分硫蛋白基因的燕麦的存在,通过扩增部分晚期胚胎发生蛋白的部分的芸豆的存在,通过部分凝集素基因的扩增的大豆的存在和通过扩增凝集素的蚕豆的存在结节蛋白基因的一部分。板栗thaumatine基因用作对照。优化PCR条件以检测1%的掺假。双重PCR和经过特殊设计的引物组可以扩增几个相关物种,从而限制了所需分析的数量和成本。结果的解释通过使用决策树来促进。

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