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A novel real-time polymerase chain reaction method for the detection of pecan nuts in food

机译:一种检测食品中山核桃果仁的实时聚合酶链反应新方法

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摘要

A real-time PCR-based method for the detection of the pecan (Carya illinoiensis) component in food is described. The method consists of DNA isolation by chaotropic solid phase extraction and the subsequent PCR with pecan-specific primers and a TaqMan fluorescent probe. The primers and the probe are targeted to the putative gene for allergenic vicilin-like seed storage protein of pecan. The method was positive for 10 pecan varieties and negative for all other tested plant materials used in food industry, including walnut. The intrinsic detection limit of the method was 1 pg pecan DNA which corresponds to 1.2 haploid genome copies. Using a series of model pastry samples with defined pecan contents, a practical detection limit of 0.01% (w/w) pecan was estimated. Practical applicability of the PCR method was tested by the analysis of 13 food samples; no discrepancies between the declared and detected pecan contents were found. The presented PCR method is useful for sensitive and selective detection of pecans in food samples and can be performed in one working day.
机译:描述了一种基于实时PCR的食品中山核桃(伊利诺伊山核桃)成分检测方法。该方法包括通过离液序列高的固相提取法分离DNA,然后使用山核桃特异的引物和TaqMan荧光探针进行PCR。引物和探针针对山核桃的变应原性丝胶样种子贮藏蛋白的推定基因。该方法对10个山核桃品种呈阳性,对食品工业中使用的所有其他测试植物材料(包括核桃)呈阴性。该方法的固有检测极限是1 pg山核桃DNA,对应于1.2个单倍体基因组拷贝。使用一系列具有确定的山核桃含量的模型糕点样品,估计实际检测极限为0.01%(w / w)山核桃。通过分析13种食品样品,检验了PCR方法的实际适用性。没有发现申报的和检测到的山核桃含量之间存在差异。提出的PCR方法可用于敏感和选择性检测食品样品中的山核桃,并且可以在一个工作日内完成。

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