Taqman-LNA荧光PCR快速检测肉制品中马源性成分的研究  

Development of a Taqman-LNA PCR Assay for Rapid Detection of Horse-Derived Ingredients in Meat Products

作　　者：许如苏[1] 周广彪[1] 段建发[1] 苏建晖[1] 魏霜[1] 陈冠武[1] 

机构地区：[1]汕头出入境检验检疫局,广东汕头515031

出　　处：《中国动物检疫》2015年第7期62-66,共5页China Animal Health Inspection

基　　金：广东检验检疫局科研课题(2014GDK24)

摘　　要：[目的]建立快速检测肉制品中马源性成分的Taqman-LNA荧光PCR方法。[方法]基于马的种属保守序列,设计特异性引物和Taqman-LNA探针,建立可快速检测肉制品中马源性成分的Taqman-LNA荧光PCR检测方法。通过对特异性、灵敏度、重复性的检测,对建立的方法进行评价。[结果]建立的Taqman-LNA荧光PCR方法灵敏,可检测到马肉DNA最低限为1.4 pg;特异,与猪、牛、羊、鹿、驴、兔、鸡、鸭、鹅、虾无非特异性反应;可重复,批内和批间的变异系数均小于2%;准确,应用该法检测人为掺入马肉的肉制品,检测结果与预期相符。[结论]本研究建立的方法具有灵敏度高,特异性强,重复性好等优点,适合于肉制品中马源性成分检测。Objective]To develop a Taqman-LNA PCR assay for rapid detection ofhorse-derived ingredients in meat products. [Method] A Taqman-LNA PCR was developed for rapid detection ofhorse-derived ingredients in meat products,using the horse-specific primers and Taqman-LNA probe designed according to the conservative domain gene sequences of horse,and evaluated for its sensibility,specificity and reproducibility. [Results] The method was sensitive with detecting limit of 1.4pghorse DNA and was specific without cross-reaction with pork,beef,mutton,deer,donkey,rabbit,chicken,duck,goose and shrimp. The repeatability test showed that the coefficient of variation of intra-assay and inter-assay was less than 2%. The method was used to detect meat products mixed with horse flesh,obtaining the expected results. [Conclusion]The established Taqman-LNA PCR possessed high sensibility,good specificity and excellent reproducibility,and was fit for rapid detection ofhorse-derived ingredients in meat products.

关 键 词：马源性成分 锁核酸(LNA)探针 荧光定量PCR 肉制品 

分 类 号：TS207.3[轻工技术与工程—食品科学]