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A Microarray-based Detection System for Genetically Modified (GM) Food Ingredients

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Abstract

A multiplex DNA microarray chip was developed for simultaneous identification of nine genetically modified organisms (GMOs), five plant species and three GMO screening elements, i.e. the 35S promoter, the nos terminator and the nptII gene. The chips also include several controls, such as that for the possible presence of CaMV. The on-chip detection was performed directly with PCR amplified products. Particular emphasis was placed on the reduction of the number of PCR reactions required and on the number of primers present per amplification tube. The targets were biotin labelled and the arrays were detected using a colorimetric methodology. Specificity was provided by specific capture probes designed for each GMO and for the common screening elements. The sensitivity of the assay was tested by experiments carried out in five different laboratories. The limit of detection was lower than 0.3% GMO for all tests and in general around 0.1% for most GMOs. The chip detection system complies with the requirements of current EU regulations and other countries where thresholds are established for the labelling of GMO.

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Abbreviations

CaMV:

cauliflower mosaic virus

CTAB:

hexadecyltrimethylammonium bromide

DNA:

deoxyribonucleic acid

EC:

European Commission

EU:

European Union

GMO:

genetically modified organism

NptII:

neomycin phospho transferase originating from Streptomyces hygroscopicus

P-35S:

promoter 35S originating from cauliflower mosaic virus

Pat:

phosphinotricin-N-acetyl transferase

PCR:

polymerase chain reaction

RRS:

roundup ready® soybean

T35S:

terminator 35S originating from cauliflower mosaic virus

Ta:

annealing temperature

T-nos:

terminator nopaline synthase originating from Agrobacterium tumefaciens

UNG:

uracil-N-glycosylase

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Authors and Affiliations

  1. Unité de Recherche en Biologie Cellulaire (URBC), Faculté Universitaire Notre Dame de la Paix, 61, rue de Bruxelles, Namur, 5000, Belgium

    Serge Leimanis, Thomas Glouden & José Remacle

  2. Molecular Genetics Department, Instituto de Biología Molecular de Barcelona (IBMB) Consejo Superior de Investigaciones Científicas (CSIC), Jordi Girona Salgado 18-26, Barcelona, 08034, Spain

    Marta Hernández, Maria Pla & Pere Puigdomènech

  3. Laboratoire de méthodologies de la détection des OGM, INRA Versailles, Route de Saint Cyr RD 10, Versailles Cedex, 78026, France

    Sophie Fernández, Francine Boyer & Yves Bertheau

  4. LGC Limited, Queens Road, Teddington, Middlesex, TW11 OLY, UK

    Malcolm Burns & Neil Harris

  5. Agency BATS, Clarastrasse 13, Base, 4058, Switzerland

    Shirin Bruderer & Othmar Kaeppeli

  6. Laboratoire Interrégional, DGCCRF, 67 Chemin du routoir, Illkirch-Graffenstaden, 67400, France

    Patrick Philipp

  7. TEPRAL—Brasseries KRONENBOURG, 68, route d’Oberhausbergen, Strasbourg, 67037, France

    Marc Vaitilingom

Authors
  1. Serge Leimanis
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  2. Marta Hernández
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  3. Sophie Fernández
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  4. Francine Boyer
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  5. Malcolm Burns
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  6. Shirin Bruderer
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  7. Thomas Glouden
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  8. Neil Harris
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  9. Othmar Kaeppeli
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  10. Patrick Philipp
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  11. Maria Pla
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  12. Pere Puigdomènech
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  13. Marc Vaitilingom
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  14. Yves Bertheau
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  15. José Remacle
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Corresponding author

Correspondence to José Remacle.

Additional information

Serge Leimanis, Marta Hernández, Sophie Fernández: These authors contributed equally to this paper.

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Leimanis, S., Hernández, M., Fernández, S. et al. A Microarray-based Detection System for Genetically Modified (GM) Food Ingredients. Plant Mol Biol 61, 123–139 (2006). https://doi.org/10.1007/s11103-005-6173-4

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  • DOI: https://doi.org/10.1007/s11103-005-6173-4

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