应用多重Taqman-LNA荧光PCR同时检测肉制品中猪鸡鸭源性成分

Development of Multiplex Taqman-LNA Real-time PCR For Detecting Pork,Chicken and Duck-Derived Ingredients in Meat Products

为了建立同时检测肉制品中猪、鸡、鸭3种动物源性成分的多重Taqman-LNA荧光PCR,给打击肉制品掺假提供技术手段。针对动物线粒体基因组的保守序列,设计特异性引物和Taqman-LNA探针,建立同时检测肉制品中猪、鸡、鸭源性成分的多重荧光PCR方法,并应用于市售肉制品的检测。建立的多重Taqman-LNA荧光PCR方法可同时检测肉制品中上述动物源性成分,与牛、羊、驴、兔、鹅等动物源性成分无交叉反应,其检测限均达到肉含量的0.001%。对170份市售肉制品检测发现,有42份样品检出与标签标示成分不符的肉种成分,样品不合格率为24.7%。本研究建立的多重Taqman-LNA荧光PCR方法可同时检测肉制品中猪、鸡、鸭3种动物源性成分,具有特异、快速、经济、高效等优点,可适应于肉制品中多种肉源性成分的高通量检测。

To develop a multiplex taqman-LNA real-time PCR for detecting pork, chicken and duck-Derived Ingredients in meatand products simultaneously. Species-specific primers and taqman-LNA probes were designed based on the COl gene sequences ofthe above animals. A multiplex taqman-LNA real-time PCR was developed for detecting the above animals-derived ingredients inmeat and products after optimization of the reaction conditions. The assay was used to detect 170 samples of meat products from mar-kets. The assay could detect 0.001% each target meat of three meat from the above animals in meat products simultaneously, and hadno cross-reaction with the DNA from cattle, sheep, donkey, rabbit, goose and other animals. 42 of 170 samples from markets werefound to have some animal-derived ingredients that were not marked on the meat product labels. The assay was rapid, specific, eco-nomic and high-throughput to detect pork, chicken and duck-derived ingredients in meat products simultaneously.