肉制品中羊源性成分的定性和定量检测

通过检测肉制品中DNA的来源进行动物源性检测是打击鲜肉及加工肉制品制假掺假的重要技术手段。依据GB/T 25165—2010《明胶中牛、羊、猪源性成分定性检测方法 实时荧光PCR法》合成用于TaqMan 实时荧光聚合酶链式反应的引物和探针，利用鲜肉及加工肉制品进行羊源性成分定性和定量检测方法研究。首先利用12 种不同动物鲜肉组织的DNA检测方法的特异性，然后以羊源DNA梯度稀释液为模板进行灵敏度实验，最后在加工肉制品中检测方法的适用性和定量检测能力。结果表明：此方法具有羊源性成分检测特异性，对其他动物来源的鲜肉DNA均无扩增信号，可以检出羊肉和猪肉混合样品中0.1%的羊肉；方法灵敏度高，可以检出10 pg的羊源DNA；方法适用性广，可以对加工肉制品（羊肉干）进行羊源性成分的定性和定量检测。

Qualitative and Quantitative Determination of Sheep-Derived Ingredients in Adulterated Meat Products

Authentication of meat products through the source identification of DNA is an important technical means for fight against meat adulteration. In this study, we developed a TaqMan-based real-time fluorescence polymerase chain reaction (RT-PCR) assay for qualitative and quantitative determination of sheep-derived ingredients in adulterated meat and meat products. Sheep-specific primers and probes were designed according to the Chinese National Standard (GB/T 25165?2010). The specificity of the PCR assay was evaluated by using it to detect DNA from 12 different animal species. Diluted sheep DNA solutions were utilized as templates to test the sensitivity of this method. Finally, its applicability and potential for quantitative determination were tested in processed meat products. The results showed high specificity for sheep rather than other animals, the ability to detect 0.1% sheep meat when mixed with pork, high sensitivity (10 pg of ovine DNA), and wide applicability to qualitatively and quantitatively identify sheep-derived ingredients in adulterated processed meat products.