Abstract
A real-time PCR-based method for construct-specific detection of the genetically modified (GM) linseed event ’CDC Triffid’ FP967 originating from Canada has been validated in a collaborative trial. The construct-specific method amplifies a 105 bp long fragment of the transgenic insertion present in FP967 spanning the junction of the terminator region of the nopalin synthase gene from Agrobacterium tumefaciens (Tnos) to a sequence region coding for the dehydrofolate reductase gene (dfr) from a class I integron from Escherichia coli. This region is characteristic for the construct used to develop FP967. A total of 11 laboratories participated in the collaborative study. For PCR analysis, each laboratory received 14 DNA samples comprising 7 double-blind DNA samples. The samples consisted of two low GM-levels of FP967 DNA (10 or 50 copies per PCR), of DNA from two different GM-positive linseed products and of DNA from GM-negative linseed, potato and rapeseed materials, respectively. All but one of the FP967-positive DNA samples were detected correctly. No false-positive results were reported. The results demonstrate that the linseed event FP967 is detectable even at low copy number concentrations. The limit of detection (LOD) determined with plasmid DNA was shown to be at 5 copies of the Tnos–dfr sequence. The data provided show that the method can be applied successfully in different laboratories and is fit-for-purpose to test for the presence of the EU-unauthorised linseed event ‘CDC Triffid’ FP967.
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Acknowledgments
The authors are very grateful to Helmut Würdemann and Beate Mühlbauer for their excellent technical assistance during this study. We would like to thank Joachim Bendiek (BVL) for carefully reading the manuscript and for helpful comments. The authors are also grateful to all participants of the collaborative trial: Sabine Burkhardt, Landeslabor Berlin Brandenburg (Berlin, Germany), Sabine Domey, Thüringer Landesanstalt für Landwirtschaft (Jena, Germany), Anette Hilpert, Thüringer Landesamt für Lebensmittel und Verbraucherschutz (Jena, Germany), Rupert Hochegger, Österreichische Agentur für Gesundheit und Ernährungssicherheit (Wien, Austria), René Köppel, Kantonales Labor (Zürich, Switzerland), Dietrich Mäde, Landesamt für Verbraucherschutz Sachsen Anhalt (Halle, Germany), Gabi Mücher, GEN-IAL GmbH (Troisdorf, Germany), Gabriele Näumann, Institut für Hygiene und Umwelt (Hamburg, Germany), Ralf Reiting, Landesbetrieb Hessisches Landeslabor (Kassel, Germany), Brigitte Speck Landwirtschaftliches Technologiezentrum (Augustenberg, Germany), Jutta Zagon, Bundesinstitut für Risikobewertung (Berlin, Germany).
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Grohmann, L., Busch, U., Pecoraro, S. et al. Collaborative trial validation of a construct-specific real-time PCR method for detection of genetically modified linseed event ‘CDC Triffid’ FP967. Eur Food Res Technol 232, 557–561 (2011). https://doi.org/10.1007/s00217-010-1403-7
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DOI: https://doi.org/10.1007/s00217-010-1403-7