锁核酸探针多重荧光PCR快速检测肉制品中4种动物肉掺假的研究  

Development of Multiplex Taqman-LNA Real-time PCR for Rapid Detecting Four Meat Adulteration in Meat and Meat Products

作　　者：许如苏[1] 周广彪[1] 魏霜[1] 段建发[1] 刘中勇[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：《检验检疫学刊》2016年第2期12-16,共5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)

摘　　要：应用锁核酸探针技术建立同时检测肉制品中牛、猪、鸡、鸭4种动物肉掺假的方法。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牛、猪、鸡、鸭源性成分的多重Taqman-LNA荧光PCR方法,用于市售肉制品的检测,同时采用标准方法进行验证。结果显示,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,检测限均达到肉含量的0.01%。将样品Ct≤30,且|样品Ct-纯肉Ct|≤7判为肉掺假;将样品Ct≤30,且|样品Ct-纯肉Ct|>7判为肉污染。对200份市售肉制品检测发现,有42份样品掺入标签标示以外肉种,有22份样品存在肉污染,其中2份样品暨掺假又污染。多重方法对肉源性成分的检测结果与现行标准方法符合率达99%。本研究建立的方法可同时检测肉制品中牛、猪、鸡、鸭4种动物源性成分,具有特异、快速、经济、高通量等优点,设定的肉掺假和肉污染判定标准合理,可有效避免肉污染导致的掺假误判。To develop a multiplex taqman-LNA chicken and duck adulteration in meat and real-time PCR for the rapid detection of bovine,pork, products simultaneously, species-specific primers and taqman-LNA probes were designed based on the CO1 gene sequence of bovine,pork,chicken and duck. A multiplex taqman-LNA real-time PCR was developed for the rapid detection of the animals-derived ingredients in meat and products after optimization of the reaction conditions. The assay and standard method were simultaneously used to detect 200 samples from markets. The results showed that the assay could detect 0.01% each target meat of four meat from the above animals in meat products simultaneously. There was positive of adulteration for sample with the result I Ct （sample） - Ct（meat） ｜ ≤7, and positive of pollution for sample with the result I Ct（sample）-Ct（meat）｜〉7 ＆ Ct（sample）≤30.42 of 200 samples from markets were found adulteration,21 of 200 meat products were found pollution by other meat. The results for detecting the samples by the assay were 99% consistent with the results of the standard methods. The assay was economic,high-throughput,rapid and specific,fit to detect bovine, pork,chicken and duck adulteration in meat and meat products simultaneously. There was a rational criterion for the result to avoid mistaking contamination to adulteration of meat effectively.

关 键 词：动物源性成分 掺假 锁核酸探针 多重Taqman-LNA荧光PCR 肉制品 

分 类 号：TS207.3[轻工技术与工程—食品科学]