摘要
[目的]建立快速检测肉制品中猪源性成分的锁核酸探针荧光PCR检测方法。[方法]基于猪线粒体基因组的保守序列,设计特异性引物和锁核酸探针,建立快速检测肉制品中猪源性成分的锁核酸荧光PCR检测方法。通过特异性、灵敏度、重复性测试及应用检测对建立的方法进行评价。[结果]建立的锁核酸荧光PCR方法对牛、羊、马、鸡、鸭、鹅均无非特异性扩增;该法可检测低至1.20 pg的猪肉DNA,重复检测结果的变异系数均小于1%。应用该法检测市售肉制品50份,32份标示含有猪肉成分样本的检测结果均与预期相符,另发现1份羊肉丸为猪肉制备、2份鸡肉火腿肠中掺有猪源性成分而未标示。[结论]本研究建立的方法具有特异、灵敏、稳定等优点,适用于大规模快速检测肉制品中猪源性成分。
Objective] To develop a LNA-qPCR method for rapid detection of pig-derived ingredients in meat and products. [Method] A LNA-qPCR method was developed for rapid detection of pig-derived in-gredients in meat and products,using the pig-specific primers and locked nucleic acid probe designed for the conservative domain gene of pig mtDNA,and evaluated by sensibility,specificity,reproducibility and Practicability. [Results] The method had no cross-reaction with the ingredients derived from beef, mutton,horse,chicken,duck and goose. It could detect pig DNA as little as 1.20 pg and it was repeat-able that the coefficients of variation were both less than 1%. The pig-derived ingredients were found in 32 meat products when detecting 50 meat products,which were in line with expectations. Also found 1 mutton pills was made of pork and 2 chicken ham sausage had pig-derived ingredients without any pig-meat in identification. [Conclusion] The LNA-qPCR established method was specific,sensitive,repeatable and accurate,and it was fit for large detecting pig-derived ingredients in meat and products rapidly.
出处
《检验检疫学刊》
2015年第4期12-15,共4页
Journal of Inspection and Quarantine
基金
广东出入境检验检疫局科技计划项目(2014GDK24)
关键词
猪源性成分
锁核酸探针
肉制品
Pig-Derived Ingredients
Locked Nucleic Acid Probe
Meat and Products
