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Development of a modular system for detection of genetically modified organisms in food based on ligation-dependent probe amplification

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Abstract

The application of a method based on ligation-dependent probe amplification to the simultaneous detection of different genetically modified organisms in food is described. The ligation reaction of target-specific probes with characteristic lengths and universal primer binding sites is followed by PCR amplification using fluorescein-labeled primers. The separation and the detection of DNA fragments are achieved by capillary electrophoresis via laser-induced fluorescence. The approach allows the simultaneous detection of several targets corresponding to different levels of specificity in a one-tube assay. Synthetic oligonucleotides were designed to detect (1) reference genes in the genome from maize, soya and rapeseed, (2) the CaMV 35S-promotor as screening element, (3) the construct-specific 35S-pat junction, and (4) the event-specific regions of the transgenic maize line MON 810 and of Roundup Ready soya. Specificity and sensitivity (GMO content 0.1%) of the approach were shown for all targets. This novel analytical strategy represents a flexible, modular system for the surveillance of GMO-derived products that can be readily complemented by further target sequences of interest.

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Authors and Affiliations

  1. Wissenschaftszentrum Weihenstephan, Technische Universität München, Lehrstuhl für Allgemeine Lebensmitteltechnologie, Am Forum 2, 85350, Freising-Weihenstephan, Germany

    Alexandra Ehlert & Karl-Heinz Engel

  2. Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit LGL, Veterinärstr. 2, 85764, Oberschleißheim, Germany

    Francisco Moreano & Ulrich Busch

Authors
  1. Alexandra Ehlert
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  2. Francisco Moreano
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  3. Ulrich Busch
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  4. Karl-Heinz Engel
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Correspondence to Karl-Heinz Engel.

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Ehlert, A., Moreano, F., Busch, U. et al. Development of a modular system for detection of genetically modified organisms in food based on ligation-dependent probe amplification. Eur Food Res Technol 227, 805–812 (2008). https://doi.org/10.1007/s00217-007-0790-x

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  • DOI: https://doi.org/10.1007/s00217-007-0790-x

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