建立并验证一种用于同时检测食品中三种芥菜类蔬菜（白芥、黑芥和芥菜）的双重实时荧光 PCR 方法。
Development and validation of a duplex real-time PCR assay for the simultaneous detection of three mustard species (Sinapis alba, Brassica nigra and Brassica juncea) 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.
Department of Analytical Chemistry, University of Vienna, Währinger Straße 38, 1090 Vienna, Austria.
出版信息
Food Chem. 2014 Jun 15;153:66-73. doi: 10.1016/j.foodchem.2013.12.035. Epub 2013 Dec 12.
The paper presents a duplex real-time PCR assay for the simultaneous detection of three potentially allergenic mustard species commonly used in food: white mustard (Sinapis alba), black mustard (Brassica nigra) and brown mustard (Brassica juncea). White mustard is detected in the "green" and black/brown mustard in the "yellow" channel. The duplex real-time PCR assay does not show cross-reactivity with other Brassicaceae species including broccoli, cauliflower, radish and rapeseed. Low cross-reactivities (difference in the Ct value ⩾ 11.91 compared with the positive control) were obtained with cumin, fenugreek, ginger, rye and turmeric. When applying 500 ng DNA per PCR tube, the duplex real-time PCR assay allowed the detection of white, black and brown mustard in brewed model sausages down to a concentration of 5mg/kg in 10 out of 10 replicates. The duplex real-time PCR assay was applied to verify correct labelling of commercial foodstuffs.
本文提出了一种双重实时 PCR 检测方法，用于同时检测三种常用于食品的潜在过敏原性芥菜属物种：白芥菜（Sinapis alba）、黑芥菜（Brassica nigra）和褐芥菜（Brassica juncea）。白芥菜在“绿色”通道中检测，黑/褐芥菜在“黄色”通道中检测。双重实时 PCR 检测方法与其他芸薹属物种（包括西兰花、花椰菜、萝卜和油菜籽）无交叉反应性。与阳性对照相比，小茴香、胡芦巴、生姜、黑麦和姜黄的 Ct 值差异 ⩾11.91，表明存在低交叉反应性。当每个 PCR 管使用 500 ng DNA 时，该双重实时 PCR 检测方法可在 10 个重复试验中的 10 个中检测到酿造模型香肠中白、黑、褐芥菜的浓度低至 5mg/kg。该双重实时 PCR 检测方法用于验证商业食品的正确标签。
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