中国农业科学2023，Vol.56Issue(22)：4371-4385,15.DOI:10.3864/j.issn.0578-1752.2023.22.002
转基因定量检测结果测量不确定度的自上而下评定方法研究及应用
Development and Application of Top-Down Approaches for Estimating Measurement Uncertainty of GMO Quantitative Results
摘要
Abstract
[Objective]The enforcement of labeling regulations on genetically modified organisms(GMOs)requires establishing a standard system for accurate quantification of GMOs that includes the standard or guide for estimating measurement uncertainty(MU).It is urgent to establish a standardized top-down approach for estimating MU of quantitative results,which is conveniently adopted by the general testing laboratories.[Method]There are two approaches for estimating the MU introduced by precision of quantitative method,one is to establish the equation of MU estimation using data obtained on 15 routine samples based on the"uncertainty function",the other is to evaluate the MU by repeatedly measuring a certified reference material(CRM)and calculating the intermediate precision.The uncertainty introduced by bias is evaluated using a CRM or a sample prepared by laboratory as bias control.The uncertainty of the nominal value of the sample prepared by laboratory is evaluated by using a simplified program based on the preparation process.The MU contributed by method precision and bias are combined into the standard uncertainty of the quantitative results,and then multiplied by the coverage factor k to obtain the expanded uncertainty.[Result]The event-specific PCR method of genetically modified maize DBN9936 was took as an example.The MU of method precision was evaluated to be 0.76%using simulated DBN9936 routine samples,and to be 0.33%using a CRM(GBW(E)100901).Compared with routine samples,the MU of method precision evaluated using a CRM is significantly underestimated.The uncertainty introduced by bias was evaluated to be 0.26%using a CRM(GBW(E)100901)as a bias control.Using a laboratory prepared powder sample and a genomic DNA sample(nominal values of 3.0%)as bias control,the bias uncertainty was evaluated to be 0.20%and 0.19%,respectively.Since the simplified program ignored some uncertainty components,the uncertainty of the nominal value of laboratory prepared samples was estimated to be smaller.By combining the MU of method precision and bias,the expanded uncertainty using routine samples was obtained to be 1.26%,1.20%,and 1.20%,respectively,the expanded uncertainty using a CRM was 0.84%,0.78%,and 0.76%,respectively.[Conclusion]This study established the top-down approaches for MU estimation of quantitative results,testing laboratories should prioritize routine samples to estimate the MU contributed by the method precision,and select CRMs as bias control in principle to evaluate bias uncertainty during GMO quantification.关键词
转基因定量结果/测量不确定度/自上而下/有证标准物质/阳性定量质控品Key words
quantitative results of GMO content/measurement uncertainty/top-down approach/certified reference materials/bias control引用本文复制引用
李俊,陈子言,吴刚,沈平,徐利群,武玉花,赵新,陈红,李飞武,梁晋刚,李允静,王颢潜,高鸿飞,张华..转基因定量检测结果测量不确定度的自上而下评定方法研究及应用[J].中国农业科学,2023,56(22):4371-4385,15.基金项目
科技创新2030—重大项目(2022ZD0402010) （2022ZD0402010）
