Characterization of Genetically Modified Canola T45 Plasmid Reference Material
DONG Lian-hua1,YU Xiao-bo1,4,SONG Gui-wen2,CAO Ying-long3, LI Liang1,WANG Jing1,LU Chang-ming3
1.National Institute of Metrology, Beijing 100013, China
2. Science and Technology Development Center, The Ministry of Agriculture, Beijing 100122, China
3. Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, Hubei 430062, China
4. College of Life Sciences, China Jiliang University, Hangzhou, Zhejiang 310018, China
Abstract：The key parameters of the real time PCR method for genetically modified canola T45 were validated , the results show that the real time PCR method can be used to co-validate the reference value for GM canola T45 reference material. Six laboratories are chosen to characterize the property values of GM T45 plasmid reference materials with the same method. The slop (PCR amplification efficiency), intercept and linearity coefficient of GM T45 genomic DNA and plasmid DNA are analyzed. The results show that the PCR amplification efficiency and linearity coefficient between genomic DNA and plasmid DNA are not significantly different. The statistic test shows the data from different labs are equal. The property value and expanded uncertainties for GM canola T45 plasmid reference material is 0.98±0.02 (k=2).
［1］GMDD.GMO Detection method database［DB］.http://gmdd.shgmo.org/crm/list,2011-11-10.
［2］Rnning S B, Vatilingom M, Berdal K G, et al.Event specific real-time quantitative PCR for genetically modified Bt11 maize (Zea mays) ［J］.Euro Food Res Technol, 2003, 216(4):347-354.
［3］Holst-Jensen A, Rnning S B, Lvseth A, et al.PCR technology for screening and quantification of genetically modified organisms (GMOs) ［J］.Anal Bioanal Chem, 2003, 375(8):985-993.
［4］Cankar K, tebih D, Dreo T, et al.Critical points of DNA quantification by real-time PCR—effects of DNA extraction method and sample matrix on quantification of genetically modified organisms ［J］.BMC Biotechnol, 2006, 14（6）:37-51.
［5］European commission.Event-specific method for the quantification of oilseed rape line T45 using real-time PCR ［R］. CRLVL14/04VP,2006.
［6］于亚东.化学分析的方法确认［M］. 北京:中国计量出版社, 2009.
［7］ENGL.Definition of minimum performance requirements for analytical methods of GMO testing［R］.JRC49650,2008
［8］JJG 1006—94, 一级标准物质技术规范 ［S］.
［9］ISO Guide 35:2006, Reference materials -general and statistical principles for certification ［S］.
2011, Vol. 32 
