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Report on the Verification of the Performance of a Construct-Specific Assay for the Detection of Flax CDC Triffid Event FP967 Using Real-Time PCR

Abstract

Further to the detection by the German authorities of the unauthorised flax CDC Triffid event FP967 (Unique Identifier CDC-FLØØ1-2) in materials imported from Canada, a notification was sent through the Rapid Alert System for Food and Feed (RASFF) in September 2009. On 21st August 2009, the Community Reference Laboratory for Genetically Modified Food and Feed received from the German authorities a construct-specific method for the detection of flax CDC Triffid event FP967, developed by Genetic ID, Augsburg (Germany). The method developer declared this method as specific for event FP967 as it targets a transition sequence spanning the nopaline synthase (nos) terminator gene and the spectinomycin/streptomycin resistance gene, construction being found only in the flax FP967 event. On 11th September 2009, the CRL-GMFF received from the German authorities the FP967 positive control in the form of DNA extracted from seeds. Seeds were provided to the German authorities by the University of California, Riverside, USA. The CRL-GMFF carried out experiments on the control sample received in order to verify the specificity and the Limit of Detection (LOD) of the construct-specific method. The CRL-GMFF observed that the NOST-Spec (nos terminator ¿ spectinomycin resistance gene) construct-specific method generates a PCR amplification product of 105 bp, whose sequence is homologous to a transition sequence spanning the nopaline synthase (nos) terminator gene and the dihydrofolate reductase gene. The experimental testing of the specificity indicates that the NOST-Spec construct-specific assay does not detect genetically modified events under the conditions reported. The limit of detection (LOD) established is between 1 and 5 haploid genome copies of FP967.JRC.DDG.I.4-Molecular biology and genomic

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