Simultaneous Detection of Eight Dairy-Derived Components Using Double-Tube Multiplex qPCR Based TaqMan Probe

文献类型： 外文期刊

第一作者： Su, Yingying

作者： Su, Yingying;Meng, Lu;Wang, Jiaqi;Zheng, Nan;Zhao, Yankun

作者机构：

关键词： double-tube and multiplex qPCR; TaqMan probes; milk and dairy products; adulteration; PCR efficiency; limit of detection

期刊名称：FOODS （ 影响因子：5.1； 五年影响因子：5.6 ）

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年卷期： 2024 年 13 卷 20 期

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收录情况： SCI

摘要： The authentication of milk and dairy products has great significance for food fraud. The present investigation entailed the development of a novel method that amalgamates the double-tube approach with multiplex real-time polymerase chain reaction (PCR) technology, incorporating TaqMan probes, to facilitate the high-throughput screening and detection of animal-derived constituents within milk and dairy products. Eight dairy-derived animal-specific primers and probes were designed for the mitochondrial cytochrome b (Cytb) gene of eight dairy products, including cow, buffalo, yak, goat, sheep, horse, donkey, and camel. Through the developed double-tube detection assays, the above eight targets could be simultaneously identified with a detection limit of 0.00128-0.0064 ng/mu L. The multiplex qPCR assay was effectively validated using simulated adulterated samples with different mixing ratios and demonstrated a detection limit of 0.1%. Upon analysis of 54 commercially available dairy products, a mislabeling rate of 33% was revealed. This method affords an efficacious means of detecting dairy product ingredients, thereby offering robust technical backing for market oversight and regulatory enforcement of milk and dairy products.

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