Abstract:To validate the key parameters of the real time PCR method for GM maize NK603,the result shows that this real time PCR method can be used to assign the value for GM maize NK603 matrix reference material.Seven laboratories are chosen to characterize the property values of three different GM content matrix reference materials by using the same method.Although different types of instruments are used by different lab,the standard curve generated by each lab is comparable which indicate that this method has a good reproducibility.The statistic test shows the data from different labs with equal.The values and expanded uncertainties for three matrix GM reference material are 0.57%±0.11%,1.21%±0.30% and 4.92%±1.80%(k=2).
[1]GMDD.GMO Detection method database[EB].http://gmdd.shgmo.org/crm/list,2011-01-25.
[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].European Food Research and Technology,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,Stebih 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]JRC.Report on the verification of the performance of a NK603 event-specific method on maize line NK603 using real-time PCR [R].CRLVL27/04VR,2008.
[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]Lee S,Park Y,Kim J,et al.Qualitative PCR method for detection of genetically modified maize lines NK603 and TC1507 [J].Agricul Chem Biotechnol,2004,47(4): 185-188.
[8]European commission.Event-specific method for the quantification of maize line NK603 using real-time PCR[R].Validation report NK603,2004.
[9]European Network of GMO Laboratories.Definition of minimum performance requirements for analytical methods of GMO testing [R].2008. http://gmo-crl.jrc.ec.europa.eu/doc/Min_Perf_Requirements_Analytical_methods.pdf.
[10]JJG 1006—94.一级标准物质技术规范 [S].
[11]ISO Guide 35:2006,Reference materials-general and statistical principles for certification [S].