Validation of a fast real-time PCR method to detect fraud and mislabeling in milk and dairy products

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• 作者：M. Di Domenico ; ^{m.didomenico@izs.it} ; M. Di Giuseppe ; J.D. Wicochea Rodrí ; guez ; C. Cammà
• 关键词：species identification ; dairy products ; isoelectric focusing ; real-time PCR
• 刊名：Journal of Dairy Science
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：100
• 期：1
• 页码：106-112
• 全文大小：383 K
• 卷排序：100

文摘

Fast real-time PCR TaqMan assays were developed and validated for species identification in dairy products. Based on the amplification of 12S rRNA and cytB partial genes of mitochondrial DNA, the methods were demonstrated to be sensitive, fast, and species-specific for Bos taurus, Ovis aries, Bubalus bubalis, and Capra hircus. The limit of detection calculated was lower than 1%, and the efficiency was reported to be higher than 96% in every assay. An internal amplification control was used to detect possible false negatives. The method was validated by means of laboratory-prepared samples mixing different species. Moreover, 18 commercial dairy samples were analyzed by both real-time PCR and isoelectric focusing, the official European Union reference method. The 4 TaqMan assays were confirmed to be a useful tool for milk and dairy product authentication.