Taqman探针荧光聚合酶链式反应实时同步鉴定动物源性食品中猪肉、鸡肉源性成分.docVIP

Taqman探针荧光聚合酶链式反应实时同步鉴定动物源性食品中猪肉、鸡肉源性成分 　　摘 要：目的：建立一种能实时同步检测动物源性食品中猪肉源性和鸡肉源性成分的Taqman探针双重荧光聚合酶链式反应（polymerase chain reaction，PCR）方法，应用于动物源性食品的成分掺假快速检验。方法：分别依据猪和鸡的种间保守基因（Cytb）序列设计、合成特异性引物及不同荧光标记（FAM、HEX）的Taqman探针，建立可同步检测动物源性食品中的猪源性和鸡源性成分的Taqman探针双重荧光PCR方法。结果：所建立的Taqman探针双重荧光PCR检测方法特异性强，仅对猪、鸡成分有扩增；灵敏度高，最低检测到猪源、鸡源DNA的含量分别为0.02、0.10 ng；抗干扰性强，当DNA混样中猪源、鸡源性成分含量在2%以上水平时，所建立的混合检测体系均能对DNA混样给出正确判断。结论：所建立的混合检测体系具有高特异性、高灵敏度，能够适用于动物源性食品中猪肉、鸡肉成分的同时快速检测。 　　关键词：双重荧光聚合酶链式反应；动物源性食品；猪源性成分；鸡源性成分 　　A Dual Fluorescent Real-Time Polymerase Chain Reaction Method for Concurrently Detecting Pork-Derived and Chicken-Derived Ingredients in Animal-Origin Foods 　　JIN Ping， JIE Li， LU Jun， CHEN Ying， DING Hongliu* 　　（Suzhou Quality Supervision and Inspection Center， Suzhou 215128， China） 　　Abstract： Objective： To establish a dual fluorescent real-time polymerase chain reaction （PCR） assay for the simultaneous detection of pork-derived and chicken-derived ingredients which enables rapid detection of adulteration in animal-origin foods. Methods： With this aim， specific primers and TaqMan probes were designed according to the interspecific conservative sequences. Results： The dual fluorescence PCR method had good specificity and could only amplify pork-derived ingredients and chicken-derived ingredients. The method had high sensitivity and could detect as low as 0.02 ng of porcine DNA and 0.10 ng of chicken DNA. It had strong anti-interference ability and could correctly detect more than 2% pork-derived and chicken-derived ingredients spiked in mixed DNA samples with other species. Conclusion： The dual fluorescent real-time PCR provides a specific and sensitive method for the detection of pork and chicken-derived ingredients in animal-origin foods. 　　Key words： dual fluorescent real-time polymerase chain reaction （RT-PCR）； animal-origin product； pork-derived ingredients； chicken-derived ingredients 　　DOI：10.15922/ki.rlyj.2016.09.004 　　中图分类号：TS207.3 文献标志码：A 文章编号：1001-8123（2016）09-0017-06