实时荧光PCR"自下而上"不确定度评定方法在转基因玉米成分测定中的应用
Development and Application of"Bottom-up"Evaluation Method for Measurement Uncertainty of Quantitative Detection by Real-time PCR in Genetically Modified Maize
龙丽坤 1武玉花 2张华 3何禹璇 1梁晋刚 3刘奕君 4马月 1赵新 5闫伟 1董立明 1王颢潜 3沈平 3徐利群 3李飞武1
作者信息
- 1. 吉林省农业科学院,长春 130033
- 2. 中国农业科学院油料作物研究所/农业农村部农业转基因生物溯源重点实验室,武汉 430062
- 3. 农业农村部科技发展中心,北京 100025
- 4. 吉林省农业科学院,长春 130033;吉林农业大学,长春 130061
- 5. 天津市农业科学院,天津 300384
- 折叠
摘要
研究中建立转基因成分实时荧光PCR定量检测方法的"自下而上"不确定度评定模型,提出利用实验室自制校准品测定的不确定度评定方法,将标准曲线参数斜率(a)、截距(b)和y(测定Ct值)作为输入量,获得测定结果的扩展不确定度.以转基因玉米CC-2定量检测定值为例,利用建立的实时荧光PCR定量检测"自下而上"的不确定度评定模型,采用自制校准品开展不确定度评定.结果表明,3个子样测定值的标准不确定度分别为17.74％、17.62％和17.38％,合成不确定度为10.15％,扩展不确定度为20.30％.按照本模型将转基因实验室间测定结果进行比对,结果表明,3个实验室测定的扩展不确定度分别为20.30％、13.53％和15.15％.
Abstract
In this study,a"bottom-up"uncertainty evaluation model of real-time PCR quantitative detection of GM was established,and an uncertainty evaluation method for quantitative determination of laboratory self-made calibrations was proposed.The slope(a),intercept(b)and y(measured Ct value)of the standard curve were used as in-put quantities to calculate the extended uncertainty of the measurement results.CC-2 genomic DNA was used as a self-made calibration sample,and the quantitative detection value of GM maize CC-2 was taken as an example to evaluate the uncertainty by established"bottom-up"uncertainty evaluation model.The standard uncertainties of three sub-samples were 17.74％,17.62％,and 17.38％,respectively;the synthetic uncertainty was 10.15％,and the extended uncertainty was 20.30％.The CC-2 real-time PCR method of three laboratories was compared between the laboratories,and the uncertainty was evaluated according to this model.The results showed that the extended uncer-tainty of the three laboratories was 20.30％,13.53％and 15.15％,respectively.
关键词
玉米/实时荧光PCR/定量测定/测量不确定度Key words
Maize/Real-time PCR/Quantification of GM/Measurement uncertainty引用本文复制引用
出版年
2024
NETL
NSTL
万方数据