Development and Application of"Bottom-up"Evaluation Method for Measurement Uncertainty of Quantitative Detection by Real-time PCR in Genetically Modified Maize
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.
MaizeReal-time PCRQuantification of GMMeasurement uncertainty
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