食品和饲料中转基因植物FMV35S启动子环介导等温扩增检测方法的建立Establishment of a Method to Detect Genetically Modified Plant FMV35S Promoter in Food and Feeds Using Loop-mediated Isothermal Amplification
邝筱珊;胡松楠;游淑珠;成晓维;王小玉;冯家望;
摘要(Abstract):
[目的]建立FMV35S基因启动子环介导等温扩增的检测方法。[方法]建立一种检测食品和饲料中FMV35S转基因成分的环介导恒温扩增(LAMP)检测方法,并利用实时浊度法对该LAMP检测方法进行灵敏度、特异性和稳定性评价。[结果]该方法的检测限为0.5%,特异性和稳定性良好。采用LAMP法与荧光PCR方法进行了实际样品对比检测,结果一致可靠。[结论]可采用LAMP法对食品和饲料中FMV35S转基因成分进行检测。
关键词(KeyWords): 食品;饲料;FMV35S基因;环介导恒温扩增;检测
基金项目(Foundation):
作者(Author): 邝筱珊;胡松楠;游淑珠;成晓维;王小玉;冯家望;
Email:
DOI: 10.13989/j.cnki.0517-6611.2014.16.004
参考文献(References):
- [1]JAMES C.Global Status of Commercialized Biotech/GM Crops:2012[R].Ithaca,NY,ISAAA Brief No 44 ISAAA,2012,
- [2]ZHOU X,XING D,TANG Y,et al.PCR-free detection of genetically modified organisms using magnetic capture technology and fluorescence crosscorrelation spectroscopy[J].PloS One,2009,4(11):8074.
- [3]ZHU D,TANG Y,XING D,et al.PCR-free quantitative detection of genetically modified organism from raw materials.An electrochemiluminescencebased bio bar code method[J].Analytical Chemistry,2008,80(10):3566-3571.
- [4]KIM J H,KIM S Y,LEE H,et al.An event-specific DNA microarray to identify genetically modified organisms in processed foods[J].Journal of Agricultural and Food Chemistry,2010,58(10):6018-6026.
- [5]NOTOMI T,OKAYAMA H,MASUBUCHI H,et al.Loop-mediated isothermal amplification of DNA[J].Nucleic Acids Research,2000,28(12):63.
- [6]AHMED M U,HASAN Q,MOSHARRAF HOSSAIN M,et al.Meat species identification based on the loop mediated isothermal amplification and electrochemical DNA sensor[J].Food Control,2010,21(5):599-605.
- [7]WANG F,JIANG L,YANG Q,et al.Rapid and Specific Detection of Escherichia coli Serogroups O26,O45,O103,O111,O121,O145,and O157 in Ground Beef,Beef Trim,and Produce by Loop-Mediated Isothermal Amplification[J].Applied and Environmental Microbiology,2012,78(8):2727-2736.
- [8]RANDHAWA G J,SINGH M,MORISSET D,et al.Loop-Mediated Isothermal Amplification:Rapid Visual and Real-Time Methods for Detection of Genetically Modified Crops[J].Journal of Agricultural and Food Chemistry,2013,61(47):11338-11346.
- [9]ZHANG J,FENG Y,HU D,et al.Rapid and Sensitive Detection of H7N9 Avian Influenza Virus by Use of Reverse Transcription-Loop-Mediated Isothermal Amplification[J].Journal of Clinical Microbiology,2013,51(11):3760-3764.
- [10]焦文强,殷相平,柳纪省.环介导等温扩增技术原理及其在检测诊断病原微生物中的应用[J].生物技术通报,2009(9):50-53.
- [11]ISO EN 21571:2005[J].Foodstuffs-Methods of analysis for the detection of genetically modified organisms and derived products-nucleic acid[S].2013.
- [12]FU S,QU G,GUO S,et al.Applications of loop-mediated isothermal DNA amplification[J].Applied Biochemistry and Biotechnology,2011,163(7):845-850.
- [13]CHEN X,WANG X,JIN N,et al.Endpoint visual detection of three genetically modified rice events by loop-mediated isothermal amplification[J].International Journal of Molecular Sciences,2012,13(11):14421-14433.
- [14]刘津,凌莉,吴少云,等.转基因大豆GTS 40-3-2品系特异性环介导等温扩增法检测方法的建立与应用[J].粮食与饲料工业,2012(12):60-63.
- [15]凌莉,刘静宇,易敏英,等.转基因玉米Bt176品系特异性环介导等温扩增检测方法的研究[J].食品工业科技,2013(3):310-313.