Development and Interlaboratory Validation of a Simple Screening Method for Genetically Modified Maize Using a ΔΔCq-Based Multiplex Real-Time PCR Assay

A number of genetically modified (GM) maize events have been developed and approved worldwide for commercial cultivation. A screening method is needed to monitor GM maize approved for commercialization in countries that mandate the labeling of foods containing a specified threshold level of GM crops. In Japan, a screening method has been implemented to monitor approved GM maize since 2001. However, the screening method currently used in Japan is time-consuming and requires generation of a calibration curve and experimental conversion factor (C-f) value. We developed a simple screening method that avoids the need for a calibration curve and C-f value. In this method, Delta C-q values between the target sequences and the endogenous gene are calculated using multiplex real-time PCR, and the Delta Delta C-q value between the analytical and control samples is used as the criterion for determining analytical samples in which the GM organism content is below the threshold level for labeling of GM crops. An interlaboratory study indicated that the method is applicable independently with at least two models of PCR instruments used in this study.