牛源性成分鉴定的引物和探针
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摘要
为研发具有自主知识产权的锡林郭勒盟牛源性检测试剂盒。以8种动物的线粒体全基因组序列为基础,通过比对分析筛选出牛特异的引物和探针组合进行荧光定量聚合酶链式反应(polymerase chain reaction,PCR)。以所设计的引物和探针为基础的牛源性成分检测具有高度的特异性(仅对牛源性样品有典型扩增曲线,对其他动物来源的样品无扩增曲线)。检测灵敏度试验表明,利用此引物和探针的检测方法具有高度的灵敏度,可以检测到1 pg级的模板DNA。牛肉加工制品中牛源性定量检测试验表明,所研发的引物和探针具有较好的定量检测能力。以上研究结果表明此引物和探针组合适用于牛源性成分的定性和定量检测。
To establish a detection kit of bovine origin which Xilin Gol League Independent Intellectual Property,the primers and probes were designed on the whole mitochondrial genome sequences from 8 animals.The result showed high specitivity,which no specific amplification for other kinds of animals except for bovine.The objective of this study was to design the primer and probe with intellectual property which can be used to detect bovine-derived materials by means of Taq Man PCR. The results inferred that the technique based on the primers and probe was specific for species identification and quantification,and was able to detect 1 pg template.This biotechnological method based on Taq Man PCR was appropriate to identify bovine-derived materials in farm products and foods.
To establish a detection kit of bovine origin which Xilin Gol League Independent Intellectual Property,the primers and probes were designed on the whole mitochondrial genome sequences from 8 animals.The result showed high specitivity,which no specific amplification for other kinds of animals except for bovine.The objective of this study was to design the primer and probe with intellectual property which can be used to detect bovine-derived materials by means of Taq Man PCR. The results inferred that the technique based on the primers and probe was specific for species identification and quantification,and was able to detect 1 pg template.This biotechnological method based on Taq Man PCR was appropriate to identify bovine-derived materials in farm products and foods.
引文
[1]田金辉.食品中动物源性成分定性鉴定方法研究[D].西安:西北农林科技大学,2011
[2]李家鹏,乔晓玲,田寒友,等.食品和饲料中动物源性成分检测技术研究进展[J].食品科学,2011,32(9):340-347
[3]魏小龙.浅谈问题肉的危害及鉴别办法[J].甘肃畜牧兽医,2017,47(8):33-34
[4]孟楠,樊振江.肉及肉制品动物源性成分鉴别技术研究进展[J].现代食品,2016(22):82-84
[5]石盼盼,李旭,吴昊,等.肉及肉制品中动物源性成分核酸检测方法研究进展[J].食品研究与开发,2016,37(10):211-214
[6]王思伟,张英杰.利用动物肌肉组织进行物种鉴定方法的研究进展[J].中国草食动物科学,2014(S1):42-45
[7]康文欣.肉制品和饲料中鸭源性成分PCR鉴别方法标准化研究[D].长春:吉林大学,2014
[8]REA S,STORANI G,MASCARO N,et al.Species identification in anchovy pastes from the market by PCR-RFLP technique[J].Food Control,2009,20(5):515-520
[9]CUSHWA W T,MEDRANO J F.Applications of the random amplified polymorphic DNA(RAPD)assay for genetic analysis of livestock species[J].Animal Biotechnology,1996,7(1):11-31
[10]陈益春,牛海力,孔保华.实时荧光定量PCR技术在肉及肉制品中的应用[J].食品工业科技,2017,38(17):324-328
[11]李宗梦,赵良娟,赵宏,等.肉及肉制品动物源性成分鉴别技术研究进展[J].食品研究与开发,2014,35(18):122-126
[12]何玮玲.应用多重PCR及实时荧光PCR方法鉴别食品中肉类成分[D].南京:南京农业大学,2013
[13]姜文灿,岳素文,江洪,等.Taq Man探针法实时荧光定量PCR的应用和研究进展[J].临床检验杂志,2015,4(1):797-805
[14]Navarro E,Serrano-H G,Castano M J,et al.real-time PCR detection chemistry[J].Clinica Chimica Acta,2015,439(439):231-250
[15]ZHANG C L,FOWLER M R,SCOTT N W,et al.A Taq Man realtime PCR system for the identification and quantification of bovine DNA in meats,milks and cheeses[J].Food Control,2007,18(9):1149-1158
[16]沙才华,汪文辉,黄海超,等.Taq Man荧光PCR法检测肉制品中牛源性成分及定量研究[J].中国动物检疫,2016,33(12):89-93
[17]范丽丽,李培,丁洪流,等.实时荧光PCR技术快速检测食品中的牛源成分[J].食品工业科技,2013,34(12):65-67
[2]李家鹏,乔晓玲,田寒友,等.食品和饲料中动物源性成分检测技术研究进展[J].食品科学,2011,32(9):340-347
[3]魏小龙.浅谈问题肉的危害及鉴别办法[J].甘肃畜牧兽医,2017,47(8):33-34
[4]孟楠,樊振江.肉及肉制品动物源性成分鉴别技术研究进展[J].现代食品,2016(22):82-84
[5]石盼盼,李旭,吴昊,等.肉及肉制品中动物源性成分核酸检测方法研究进展[J].食品研究与开发,2016,37(10):211-214
[6]王思伟,张英杰.利用动物肌肉组织进行物种鉴定方法的研究进展[J].中国草食动物科学,2014(S1):42-45
[7]康文欣.肉制品和饲料中鸭源性成分PCR鉴别方法标准化研究[D].长春:吉林大学,2014
[8]REA S,STORANI G,MASCARO N,et al.Species identification in anchovy pastes from the market by PCR-RFLP technique[J].Food Control,2009,20(5):515-520
[9]CUSHWA W T,MEDRANO J F.Applications of the random amplified polymorphic DNA(RAPD)assay for genetic analysis of livestock species[J].Animal Biotechnology,1996,7(1):11-31
[10]陈益春,牛海力,孔保华.实时荧光定量PCR技术在肉及肉制品中的应用[J].食品工业科技,2017,38(17):324-328
[11]李宗梦,赵良娟,赵宏,等.肉及肉制品动物源性成分鉴别技术研究进展[J].食品研究与开发,2014,35(18):122-126
[12]何玮玲.应用多重PCR及实时荧光PCR方法鉴别食品中肉类成分[D].南京:南京农业大学,2013
[13]姜文灿,岳素文,江洪,等.Taq Man探针法实时荧光定量PCR的应用和研究进展[J].临床检验杂志,2015,4(1):797-805
[14]Navarro E,Serrano-H G,Castano M J,et al.real-time PCR detection chemistry[J].Clinica Chimica Acta,2015,439(439):231-250
[15]ZHANG C L,FOWLER M R,SCOTT N W,et al.A Taq Man realtime PCR system for the identification and quantification of bovine DNA in meats,milks and cheeses[J].Food Control,2007,18(9):1149-1158
[16]沙才华,汪文辉,黄海超,等.Taq Man荧光PCR法检测肉制品中牛源性成分及定量研究[J].中国动物检疫,2016,33(12):89-93
[17]范丽丽,李培,丁洪流,等.实时荧光PCR技术快速检测食品中的牛源成分[J].食品工业科技,2013,34(12):65-67