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

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• 作者：Monika Palle-Reisch ; Margit Cichna-Markl ; Rupert Hochegger
• 关键词：Mustard ; Food allergen ; Real-time PCR ; Multiplexing ; Sinapis alba ; Brassica nigra ; Brassica juncea
• 刊名：Food Chemistry
• 出版年：15 June, 2014
• 年：2014
• 卷：153
• 期：Complete
• 页码：66-73
• 全文大小：488 K

文摘

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 鈥済reen鈥?and black/brown mustard in the 鈥測ellow鈥?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 猢?1.91 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 5 mg/kg in 10 out of 10 replicates. The duplex real-time PCR assay was applied to verify correct labelling of commercial foodstuffs.