Verification of the Performance of Bt11 and MIR604 Maize Event-specific Methods on the Maize Event Bt11 x MIR604 Using Real-time PCR. Validation Report and Protocols

Abstract

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 two quantitative event-specific methods on the maize event Bt11 x MIR604 (unique identifier SYN-BTØ11-1 x SYN-IR6Ø4-5) which combines the Bt11 and MIR604 transformation events. The two 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 maize (NP2276Bt11/NP2391MIR604), genomic DNA extracted from homogenised seeds of non-GM maize (NP2276/NP2391) and flour ground from seeds of NP2276Bt11/NP2391MIR604 and from seed of NP2276/NP2391. The EURL-GMFF prepared the 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 verification study are made publicly available at http://gmocrl. jrc.ec.europa.eu/.