采用蒙特卡洛法评定转基因水稻样品中NOS终止子的测量不确定度
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摘要
采用蒙特卡洛方法(MCM)实施"概率分布传播"来评定转基因水稻样品中NOS终止子的测量不确定度,解析转基因成分测量不确定度的概率分布。评定结果表明:NOS终止子的相对含量为2. 95%,非常接近理论含量(3%),而且其标准不确定度为2. 13×10-4,远小于1. 00×10-2。在95%的包含概率下,NOS终止子的相对含量分布在2. 91%～3. 00%非常窄的包含区间内,充分说明测量质量好,测量结果可靠; NOS终止子相对含量的概率分布呈标准正态分布,揭示转基因成分测量条件满足GUM法的假设,MCM和GUM法都可以应用于转基因成分测量不确定度评定。
Owing to the complex process of GMO testing such as multi-stages with several intermediate inputs/outputs and their joint probability distribution,the propagation of probability distribution was employed here for the first time to evaluate the MU in NOS terminator from genetically modified rice using Monte Carlo Method. The results showed that the relative content of NOS termination was 2. 95%,very close to the theoretical content( 3%),and the standard uncertainty was 2. 13 × 10-4,which was much less than 1. 00 × 10-2. In the coverage probability 95%,the relative content of NOS terminator is within a very narrow range of 2. 91% ～ 3. 00%,indicating the measurement quality in this work is very good,and the measurement results are very reliable. Moreover,the relative contents of NOS terminator followed standard normal distribution,revealing the measuring conditions in the testing of GMOs are agreement with the assumption in GUM approach. Consequently,both MCM and GUM method can be applied to evaluation of measurement uncertainty in the detection of GMOs.
Owing to the complex process of GMO testing such as multi-stages with several intermediate inputs/outputs and their joint probability distribution,the propagation of probability distribution was employed here for the first time to evaluate the MU in NOS terminator from genetically modified rice using Monte Carlo Method. The results showed that the relative content of NOS termination was 2. 95%,very close to the theoretical content( 3%),and the standard uncertainty was 2. 13 × 10-4,which was much less than 1. 00 × 10-2. In the coverage probability 95%,the relative content of NOS terminator is within a very narrow range of 2. 91% ～ 3. 00%,indicating the measurement quality in this work is very good,and the measurement results are very reliable. Moreover,the relative contents of NOS terminator followed standard normal distribution,revealing the measuring conditions in the testing of GMOs are agreement with the assumption in GUM approach. Consequently,both MCM and GUM method can be applied to evaluation of measurement uncertainty in the detection of GMOs.
引文
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[21] MacarthurR,Feinberg M,Bertheau Y. Construction of measurement uncertainty profiles for quantitative analysis of genetically modified organisms based on interlaboratory validation data[J]. Journal of AOAC international,2010,93(3):1046-1056.
[22] Lee S,Choi H,Kim E,et al. Estimation of the measurement uncertainty by the Bottom-Up approach for the determination of methamphetamine and amphetamine in urine[J]. Journal of analytical toxicology,2010,34(4):222-228.
[23] Sklerov J H,Couper F J. Calculation and Verification of Blood Ethanol Measurement Uncertainty for Headspace Gas Chromatography[J]. Journal of Analytical Toxicology,2011,35(7):402-410.
[24] Gullberg R G. Estimating the Measurement Uncertainty in Forensic Blood Alcohol Analysis[J]. Accreditation&Quality Assurance, 2006, 11(11):562-568.
[2] Tu J,Zhang G A,Datta K,et al. Field performance of transgenic elite commercial hybrid rice expressing Bacillus thuringiensisδ-endotoxin[J]. Nature Biotechnology,2000,18(10):1101-1104.
[3] Tu J, Datta K, Oliva N, et al. Site-independently integrated transgenes in the elite restorer rice line Minghui63 allow removal of a selectable marker from the gene of interest by self-segregation[J]. Plant Biotechnology Journal,2003,1(3):155-165.
[4] Wu G,Wu Y H,Nie S J,et al. Real-time PCR method for detection of the transgenic rice event TT51-1[J].Food Chemistry,2010,119(1):417-422.
[5] Zhang F L,Song J,Niu B,et al. An event-specific qualitative and real-time PCR detection of 98140 maize in mixed samples[J]. Food control,2015,57:1-8.
[6] Williams A. Measurement uncertainty in analytical chemistry[J]. Accreditation and Quality Assurance,1996,1(1):14-17.
[7] BIPM. JCGM 100:2008 Guide to the Expression of Uncertainty in Measurement[S]. 2008.
[8]刘建坤,朱家平,郑荣华.测量不确定度研究现状及进展[J].现代科学仪器,2013,(5):12-17.Liu J K,Zhu J P,Zheng R H. Research Status and Development of Evaluation of Measurement Uncertainty[J]. Modern Scientific Instruments,2013,(5):12-17.
[9] Hund E,Massar D L,Smeyers-Verbeke J. Operational definitions of uncertainty[J]. Trac-trends in Analytical Chemistry,2001,20(8):394-406.
[10]胡红波,孙桥,杜磊,等. GUM与基于观测方程的测量不确定度评估[J].计量学报,2017,38(5):656-660.Hu H B,Sun Q,Du L,et al. GUM and Analysis of Measurement Uncertainty Evaluation Using Observation Equation[J]. Acta Metrologica Sinica,2017,38(5):656-660.
[11]宋君,雷绍荣,刘勇,等.实时荧光定量PCR检测转基因玉米NK603结构特异基因的测量不确定度研究[J].西南农业学报,2012,25(4):147-151.Song J, Lei S R, Liu Y, et al. Uncertainties in measuring construct specific fragments of genetically modified maize NK603 by quantitative real time PCR[J]. Southwest China Journal of Agricultural Sciences,2012,25(4):147-151.
[12]宋君,雷绍荣,郭灵安,等.实时荧光定量PCR检测转基因玉米MON863的测量不确定度分析[J].玉米科学,2012,20(5):45-49.Song J,Lei S R, Guo L A, et al. Uncertainty in measuring flanking fragments of genetically modified maize MON863 by quantitative real time PCR[J].Journal of Maize Sciences,2012,20(5):45-49.
[13] Sega M,Pennecchi F,Rinaldi S,et al. Uncertainty evaluation for the quantification of low masses of benzo[a]pyrene:Comparison between the Law of Propagation of Uncertainty and the Monte Carlo method[J].Analytica Chimica Acta,2016,920:10-17.
[14]崔孝海,曲璐.蒙特卡罗方法在微波功率测量不确定度分析中的应用[J].计量学报,2008,29(1):77Cui X H,Qu L. The application to uncertainty analysis of microwave power measurement with Monte Carlo method[J]. Acta Metrologica Sinica,2008,29(1):77-79.
[15]梁志国,李新良,孟晓风.激光测振仪独立线性度的测量不确定度[J].计量学报,2008,29(3):242-247.Liang Z G, Li X L, Meng X F. The uncertainty evaluation of linearity of laser doppler velocimeters[J].Acta Metrologica Sinica,2008,29(3):242-247.
[16]崔伟群.利用蒙特卡罗对偶变量的方差减小技术进行B类标准不确定评定[J].计量与测试技术,2011,38(12):59-60.Cui W Q. Antithetic variables techniques to reduce the variance of Type B standard uncertainty in Monte Carlo method[J]. Metrology&Measurement Technique,2011,38(12):59-60.
[17]中华人民共和国农业部.转基因植物及产品成分检测抗虫转Bt基因水稻定性PCR方法:农业部953号公告-6-2007[S].北京:中国农业出版社,2007.
[18] Arumuganathan K,Earle E. Nuclear DNA content of some important plant species[J]. Plant Molecular Biology Reporter,1991,9(3):208-218.
[19] BIPM. JCGM 101:2008 Evaluation of measurement data—Supplement 1 to the “Guide to the expression of uncertainty in measurement”—Propagation of distributions using a Monte Carlo method[S]. 2008.
[20] Zel J,Gruden K,Cankar K,et al. Calculation of measurement uncertainty in quantitative analysis of genetically modified organisms using intermediate precision-a practical approach[J]. Journal of AOAC international,2007,90(2):582-586.
[21] MacarthurR,Feinberg M,Bertheau Y. Construction of measurement uncertainty profiles for quantitative analysis of genetically modified organisms based on interlaboratory validation data[J]. Journal of AOAC international,2010,93(3):1046-1056.
[22] Lee S,Choi H,Kim E,et al. Estimation of the measurement uncertainty by the Bottom-Up approach for the determination of methamphetamine and amphetamine in urine[J]. Journal of analytical toxicology,2010,34(4):222-228.
[23] Sklerov J H,Couper F J. Calculation and Verification of Blood Ethanol Measurement Uncertainty for Headspace Gas Chromatography[J]. Journal of Analytical Toxicology,2011,35(7):402-410.
[24] Gullberg R G. Estimating the Measurement Uncertainty in Forensic Blood Alcohol Analysis[J]. Accreditation&Quality Assurance, 2006, 11(11):562-568.
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