实时荧光定量PCR检测转基因玉米MON863结构特异基因的测量不确定度

Uncertainty in Measuring Construct Specific Fragments of Genetically Modified Maize MON863 by Quantitative Real Time PCR

[目的]对实时荧光定量PCR检测转基因玉米MON863结构特异基因的测量不确定度进行评定。[方法]使用转基因玉米MON863结构特异实时定量PCR方法测定含量为1%的转基因玉米MON863样品(CRM)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,评定测量的不确定度。[结果]研究得到uA=1.7×10-2,uB=9.0×10-4,uC=1.7×10-2,U95=0.036,测量结果为1.08%±0.036。[结论]转基因玉米MON863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。

[Objective] The study aimed at detect the uncertainty in measuring the construct specific fragments of genetically modified maize MON863 by quantitative real time PCR.[Method] The construct specific fragments in MON863 sample was determined by quantitative real time PCR,and then the uncertainty in result was evaluated according to the sources of uncertainty,the PCR system,data analysis and micropipette.[Result] A type of uncertainty was 1.7×10-2,B type of uncertainty was 9.0×10-4,combined standard uncertainty was 1.7×10-2,expanded uncertainty（U95） was 0.036 and the finally measured result was 1.08%±0.036.[Conclusion] Te main uncertainty of the result measured by this method came from the randomizing effect in the experimental process.