| 324 | 3 | 67 |
| 下载次数 | 被引频次 | 阅读次数 |
为了建立牦牛肉源性检测的实时荧光PCR方法本研究基于比较牦牛与黄牛、鸡、猪、羊、鸭等动物的Cytb基因序列差异,设计其特异性引物。特异性实验结果表明,猪肉、鸡肉和黄牛肉的DNA均不与引物发生交叉反应;灵敏度实验表明方法的最低检出限可达0.004 ng/μL;抗干扰实验结果表明,当牦牛肉的添加量为1%时仍可检出;对市场上的20份不同种肉样进行检测除牦牛肉样品检测结果呈现阳性外,其余肉样均为阴性。本实验所建立的实时荧光PCR方法为牦牛源性成分鉴别提供一种快速、准确、专属性强的检测方法。
Abstract：In order to establish a real-time PCR method for detection of yak beef origin,the specific primers are designed based on comparing the sequence differences of Cytb gene between yak cattle,chickens,pigs,sheep,and duck.Specificity test shows that the DNA of pork,chicken and yellow beef do not have a cross react with the primers;sensitivity test shows that the minimum detection limit of the method is 0.004 ng/μL;anti-interference experiment shows that it can still be detected when the addition of yak is 1%.20 different meat samples on the market are tested.Except yak meat samples show positive results,other meat samples are negative.The real-time PCR method established in this experiment provides a rapid,accurate and specific detection method for identification of yak derived components.
Chen D.,Bai F.,Zhou M.L.,Zhang X.Y.,and Wu D.J.,2008,Differentiation of Bos grunniens,Bos Taurus,and Bubalus from meat products mixture based on mitochondrion 12Sr RNA gene,Yichuan(Hereditas),30(8):1008-1014(陈冬,柏凡,周明亮,张翔宇,吴登俊,2008,基于线粒体12Sr RNA基因的PCR-RFLP鉴别混合牛肉及制品的牛种来源,遗传,30(8):1008-1014)
Doosti A.,Ghasemi Dehkordi P.,and Rahimi E.,2014,Molecular assay to fraud identification of meat products,J.Food Sci.Technol.,51(1):148-152
Gao J.,Wei D.,Zhang G.R.,and Nie L.Y.,2014,Recent progress on commonly used techniques for identification of meat species,Shipin Kexue(Food Science),35(11):356-360(高敬,魏迪,张癸荣,聂凌云,2014,常见肉类鉴别技术研究进展,食品科学,35(11):356-360)
Hafsa A.B.,Nabi N.,Zellama M.S.,Said K.,and Chaouachi M.,2016,A new specific reference gene based on growth hormone gene(GH1) used for detection and relative quantifica tion of Aquadvantage誖GM salmon(Salmo salar L.) in food products,Food Chem.,190:1040-1045
He W.L.,2013,A multiplex PCR and a taqman realtime PCR for the identification of meat species in meat products,Thesis for M.S.,College of Food Science and Technology,Nanjing Agricultural University,Supervisor:Huang M.,pp.2-5(何玮玲,2013,应用多重PCR及实时荧光PCR方法鉴别食品中肉类成分,硕士学位论文,南京农业大学食品科学技术学院,导师:黄明,pp.2-5)
Hou B.,Meng X.R.,Zhang L.Y.,Guo J.Y.,Li S.W.,and Jin H.,2015,Development of a sensitive and specific multiplex PCR method for the simultaneous detection of chicken,duck and goose DNA in meat products,Meat Sci.,101:90-94
Iwobi A.,Sebah D.,Kraemer I.,Losher C.,Fischer G.,Busch U.,and Huber I.,2015,A multiplex real-time PCR method for the quantification of beef and pork fractions in minced meat,Food Chem.,169:305-313
Ma Y.S.L.,LüM.X.,Tang J.N.,Chen J.,Zhang R.,and Long H.,2016,Meat identification based on Cytb gene sequences for the market meat products,Zhongguo Shipin Weisheng Zazhi(Chinese Journal of Food Hygiene),28(1):48-51(马伊萨兰,吕明星,唐俊妮,陈娟,张荣,龙虎,2016,用Cytb基因序列分析鉴定肉制品掺假,中国食品卫生杂志,28(1):48-51)
Ma Y.Z.,Ma D.,Bai D.S.,Li H.Y.,and Xia C.X.,2012,Recent advances in immunological detection of meat adulteration,Roulei Yanjiu(Meat Research),26(9):26-29(马永征,马冬,白娣斯,李海运,夏晨溪,2012,免疫学检测肉类制品掺假研究进展,肉类研究,26(9):26-29)
Qiu X.,Zhang L.,Wen Y.L.,Wang J.W.,Liu L.S.,Ma L.,Wu X.Z.,and Jin J.R.,2010,Nutritional composition analysis of meat from yak and yellow cattle in Sichuan,Shipin Kexue(Food Science),31(15):112-116(邱翔,张磊,文勇立,王建文,刘鲁蜀,马力,吴贤智,金洁茹,2010,四川牦牛,黄牛主要品种肉的营养成分分析,食品科学,31(15):112-116)
Thanakiatkrai P.,and Kitpipit T.,2017,Meat species identification by two direct-triplex real-time PCR assays using low resolution melting,Food Chem.,233:144-150
Velioglu H.M.,Sezer B.,Bilge G.,Baytur S.E.,and Boyaci I.H.,2018,Identification of offal adulteration in beef by laser induced breakdown spectroscopy(LIBS),Meat Sci.,138:28-33
Xu R.S.,Wei S.,Zhou G.B.,Ren J.,Liu Z.Y.,Tang S.Z.,Cheung P.C.K.,and Wu X.Y.,2018,Multiplex Taq Man locked nucleic acid real-time PCR for the differential identification of various meat and meat products,Meat Sci.,137:41-46
Yang H.,Wang X.F.,Xiao Y.P.,Wei W.,and Xu J.F.,2017,Rapid detection of chicken,duck and pork blending in beef products by multiple digital PCR,Zhejiang Nongye Xuebao(Acta Agriculturae Zhejiangensis),29(6):994-1000(杨华,汪小福,肖英平,魏巍,徐俊锋,2017,牛肉及其制品中掺入鸡肉、鸭肉和猪肉的多重数字PCR快速检测方法研究,浙江农业学报,29(6):994-1000)
Yi G.,GrabezV.,Bjelanovic M.,Slinde E.,Olsen K.,Langsrud O.,Phung V.T.,Haug A.,Oostindjer M.,and Egelandsdal B.,2015,Lipid oxidation in minced beef meat with added Krebs cycle substrates to stabilise colour,Food Chem.,187:563-571
基本信息:
DOI：10.13417/j.gab.039.004426
中图分类号:TS251.52
引用信息:
[1]郭华麟,明玥,李轲,等.基于实时荧光PCR方法的牦牛肉源性成分鉴别[J].基因组学与应用生物学,2020,39(10):4426-4431.DOI:10.13417/j.gab.039.004426.
基金信息:
四川农业大学科研兴趣项目(2018382);; 国家质检总局科技计划项目(2016IK114)共同资助
2019-04-28
2019-04-28
2019-04-28