Report on the verification of the performance of Bt11, MIR604 and GA21 maize event-specific methods on the maize event Bt11 x MIR604 x GA21 using real-time PCR : validation report and protocols

The European Union Reference Laboratory for GM Food and Feed (EURL-GMFF), established by Regulation (EC) No 1829/2003, has carried out a verification study to assess the performance of three quantitative eventspecific methods on the maize event Bt11 x MIR604 x GA21 (unique identifier SYN BTØ11-1 x SYN-IR6Ø4-5 x MON-ØØØ21-9) which combines the Bt11, MIR604 and GA21 transformation events. The three methods have been validated individually on single trait events, to detect and quantify each event in maize samples. This study was conducted according to internationally accepted guidelines (1, 2). In accordance to Regulation (EC) No 1829/2003 of 22 September 2003 on genetically modified food and feed and to Regulation (EC) No 641/2004 of 6 April 2004 on detailed rules for the implementation of Regulation (EC) No 1829/2003, Syngenta Seeds S.A.S. provided the detection methods and the control samples: genomic DNA extracted from homogenised seeds of Bt11 x MIR604 x GA21 maize (NP2673GA21xNP2171Bt11+MIR604), genomic DNA extracted from homogenised seeds of non-GM maize (NP2673/NP2171) and flour ground from seeds of NP2673GA21xNP2171Bt11+MIR604 and from seeds of NP2673/NP2171. The EURL-GMFF prepared the inhouse verification samples (calibration samples and blind samples at different GM percentages). The results of the in-house verification study were evaluated with reference to the European Network of GMO Laboratories (ENGL) method performance requirements (http://gmocrl.jrc.ec.europa.eu/guidancedocs.htm) and to the validation results on the individual parental events (http://gmo-crl.jrc.ec.europa.eu/statusofdoss.htm). The results of this EURL-GMFF in-house verification study are made publicly available at http://gmo-crl.jrc.ec.europa.eu/.