建立并验证三重实时荧光 PCR 方法，用于同时检测食品中三种芥菜物种和三种芹菜品种。
Development and validation of a triplex real-time PCR assay for the simultaneous detection of three mustard species and three celery varieties in food.
机构信息
Department of Analytical Chemistry, University of Vienna, Währinger Straße 38, 1090 Vienna, Austria; AGES, Austrian Agency for Health and Food Safety, Institute for Food Safety, Department of Molecular Biology and Microbiology, Spargelfeldstraße 191, 1220 Vienna, Austria.
AGES, Austrian Agency for Health and Food Safety, Institute for Food Safety, Department of Molecular Biology and Microbiology, Spargelfeldstraße 191, 1220 Vienna, Austria.
出版信息
Food Chem. 2015 Oct 1;184:46-56. doi: 10.1016/j.foodchem.2015.03.049. Epub 2015 Mar 20.
The paper presents a triplex real-time PCR assay allowing the simultaneous detection of three mustard species (white, black and brown mustard) and three celery varieties (celery roots, celery stalks and leaf celery) in foodstuffs. The triplex assay does not show cross-reactivity with other Brassicaceae. Low cross-reactivities were observed with fenugreek, cumin, ginger, caraway, turmeric, lovage and rye, the ΔCt values were, however, ⩾ 12 compared to positive controls. The triplex assay allows the detection of traces of DNA of the allergenic components in spite of an excess of the other DNA templates. Analysis of extracts from model sausages containing defined concentrations of mustard and celery showed that the triplex assay is applicable to both raw and processed foods. It was found to allow the detection of 1 ppm black/brown mustard and 50 ppm white mustard and celery in raw and brewed sausages with a probability ⩾ 95%.
本文提出了一种三重实时 PCR 检测方法，可同时检测食品中的三种芥菜（白芥菜、黑芥菜和棕芥菜）和三种芹菜（芹菜根、芹菜茎和叶用芹菜）。三重检测方法与其他芸薹属蔬菜没有交叉反应。与葫芦巴、孜然、生姜、葛缕子、姜黄、独活草和黑麦的交叉反应性较低，然而与阳性对照相比，ΔCt 值 ⩾ 12。尽管存在其他 DNA 模板的过剩，但三重检测方法允许检测过敏原成分的 DNA 痕迹。对含有确定浓度芥菜和芹菜的模型香肠提取物的分析表明，三重检测方法适用于生的和加工的食品。结果发现，三重检测方法能够以 ⩾ 95%的概率检测到生香肠和煮香肠中 1 ppm 的黑/棕芥菜和 50 ppm 的白芥菜和芹菜。
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