6种肉类成分多重PCR鉴别方法的建立及应用

Establishment and application of multiple PCR identification method for 6 kinds of meat ingredients

中文关键词:  多重PCR  肉类掺假  动物源成分

英文关键词:multiplex PCR  meat adulteration  animal-derived ingredients

中文摘要:

      目的 建立单管同步快速鉴别水牛、黄牛、牦牛、山羊、驴和鸭的物种成分的多重PCR鉴别方法。方法 对水牛、黄牛、牦牛、山羊、驴和鸭的线粒体全基因组, 采用Primer-blast在线软件, 设计6对特异性引物, 优化反应条件, 建立PCR体系。结果 该方法仅对水牛、黄牛、牦牛、山羊、驴和鸭肉有特异性扩增, 与其他动物的核酸无交叉反应; 同时检测6种物种成分的检测低限为10?4 ng/μL。在鸭肉中分别模拟掺入牛肉、羊肉和驴肉成分, 方法对鸭肉中掺入牛成分、羊成分和驴成分中的检出限均小于0.5%。结论 该方法特异性好、敏感度高, 可以被应用于水牛、牦牛、黄牛、山羊、驴和鸭的物种成分鉴别。

英文摘要:

      Objective To establish a single tube synchronous and rapid multipcr for identifying species components of buffalo, yellow cattle, yak, goat, donkey and duck. Methods Six pairs of specific primers were designed using Primer-blast online software for the complete genome of mitochondria of buffalo, yellow cattle, yak, goat, donkey and duck to optimize the reaction conditions and establish the PCR system. Results The method only amplified the nucleic acid of buffalo, yellow cattle, yak, goat, donkey and duck, and had no cross reaction with other animals. The limit of detection of 6 species was 10?4 ng/μL. In duck meat, beef, mutton and donkey ingredients were simulated respectively, and the limit of detection of beef, lamb and donkey ingredients in duck meat was less than 0.5%. Conclusion This method has good specificity and high sensitivity, and can be applied to the species composition identification of buffalo, yak, yellow cattle, goat, donkey and duck in the ingredient identification of above animal meat.