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Collaborative trial validation of cry1Ab/Ac and Pubi-cry TaqMan-based real-time PCR assays for detection of DNA derived from genetically modified Bt plant products

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Abstract

Presence of genetic modifications in rice products originating from China and imported to the European Union market is detected since 2006. Neither these products from China nor any other genetically modified rice lines are approved as food or feed in the EU. The transgenic rice varieties identified contain genetic elements and constructs coding for insect-resistance genes from Bacillus thuringiensis (Bt) coding for insecticidal crystal (cry) proteins. In particular, DNA sequences coding for codon-optimised or fused cry1Ab/c genes and constructs driven by the maize ubiquitin promoter (P-ubiZM1) were identified. For improved screening and identification of genetic modifications present in Asian rice products, two TaqMan-based real-time PCR assays targeting codon-optimised cry1Ab/Ac and the Pubi-cry construct have been developed. These assays have been validated in an international collaborative trial with 17 participants from 10 countries. Based on a new mathematical–statistical model and an adjusted experimental set-up of the collaborative trial, a close examination of the limit of detection (LOD 95%) and the probability of detection of the qualitative PCR assays was conducted. The evaluation of the method performance characteristics and results of the collaborative trial validation are presented.

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Acknowledgments

The authors are very grateful to Christine Degner (Landesamt für Verbraucherschutz, Halle), Sigrid Niendorf (Landesamt für Verbraucherschutz, Halle) and Franziska Duda (Hessisches Landeslabor, Kassel) for their excellent technical assistance during this study. We would like to thank Joachim Bendiek (BVL, Berlin) for carefully reading the manuscript and for helpful comments. The authors are grateful to the participants of the collaborative trial, namely Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit (Oberschleißheim, Germany); Bundesamt für Gesundheit (Bern, Switzerland); Chemisches und Veterinäruntersuchungsamt (Freiburg, Germany); Chemisches und Veterinäruntersuchungsamt Münsterland Emscher-Lippe (Münster, Germany); Chinese Academy of Inspection and Quarantine, Agro-Product Safety Research Center (Bejing, China); Service Commun des Laboratoires (Illkirch, France); Eurofins GeneScan Inc. (Metairie, USA); Genetic ID (Augsburg, Germany); Hessisches Landeslabor (Kassel, Germany); Institut für Hygiene (Hamburg, Germany); Joint Research Centre (Ispra, Italy); Kantonales Labor (Zürich, Switzerland); Laboratoire national de Santé (Luxembourg, Luxembourg); Landesamtes für Verbraucherschutz und Lebensmittelsicherheit (Braunschweig, Germany); National Research Centre on DNA Fingerprinting National Bureau of Plant Genetic Resources (New Dehli, India); Österreichische Agentur für Gesundheit und Ernährungssicherheit (Wien, Austria); and RIKILT Wageningen UR (Wageningen, the Netherlands).

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Authors and Affiliations

  1. Federal Office of Consumer Protection and Food Safety, Mauerstr. 39-42, 10117, Berlin, Germany

    Lutz Grohmann & Katrin Zur

  2. Hessian State Laboratory, Druseltalstr.67, 34131, Kassel, Germany

    Ralf Reiting

  3. State Office for Consumer Protection, Freiimfelder Str. 68, 06112, Halle (Saale), Germany

    Dietrich Mäde

  4. QuoData GmbH, Prellerstr. 14, 01309, Dresden, Germany

    Steffen Uhlig, Kirsten Simon & Kirstin Frost

  5. Genomic Resources Division, National Bureau of Plant Genetic Resources, Pusa Campus, New Delhi, 110012, India

    Gurinder Jit Randhawa

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  1. Lutz Grohmann
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  2. Ralf Reiting
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  3. Dietrich Mäde
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  4. Steffen Uhlig
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  7. Gurinder Jit Randhawa
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  8. Katrin Zur
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Correspondence to Lutz Grohmann.

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Grohmann, L., Reiting, R., Mäde, D. et al. Collaborative trial validation of cry1Ab/Ac and Pubi-cry TaqMan-based real-time PCR assays for detection of DNA derived from genetically modified Bt plant products. Accred Qual Assur 20, 85–96 (2015). https://doi.org/10.1007/s00769-015-1108-5

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