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

Uncertainty in Measuring Construct Specific Fragments of Genetically Modified Maize,Event NK603,by Quantitative Real Time PCR

应用四川省农业科学院分析测试中心实验室建立的转基因玉米NK603结构特异实时定量PCR方法测定含量为2%的转基因玉米NK603样品中结构特异片段的含量,并从扩增反应、数据处理以及微量移液器等不确定度来源,评定了测量的不确定度。结果表明,uA=0.0003,uB=0.001,uC=0.001,U95=0.002,测量结果为1.6%±0.002%。NK603结构特异实时定量PCR方法检测结果的主要不确定度来自检验过程中的随机效应。

The detection method of genetically modified maize,Event NK603,established by ourselves was used to measuring the construct specific fragment in samples and we evaluated the uncertainty of detection according to the sources of uncertainty,the PCR system,data analysis and micropipette.The results showed that A type of uncertainty was 0.0003,B type of uncertainty was 0.001,combined standard uncertainty was 0.001,expanded uncertainty（U95） was 0.002 and the finally measured result was 1.6%±0.002.Thus the main uncertainty of the result measured by this method came from the randomizing effect in the experimental process.