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Comparison of plasmid and genomic DNA calibrants for the quantification of genetically modified ingredients

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Abstract

Real-time PCR is the method of choice for the quantification of the genetically modified (GM) content of food. Recent EU Commission recommendations advocate expressing the GM content as the percentage of the number of GM target DNA sequences per target taxon specific sequence. The provision of reference materials certified for their copy number content, for example in the form of plasmids, are hence desirable to fulfil this EU recommendation. An inter-laboratory trial was conducted to compare plasmid DNA to the more traditional genomic DNA approach for quantitative PCR calibration in terms of copy numbers, using the model system of Roundup Ready™ soya. Data was analysed from 48 randomised real-time quantitative PCR plates under conditions of reproducibility across three international laboratories. Results demonstrated that the specific plasmid DNA used in the laboratory trial provided a suitable alternative to genomic DNA for use as a calibrant in GM quantification. In the current investigation, plasmid calibrants gave equal or better performance characteristics in terms of precision and closeness to the expected value, than their genomic equivalents.

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Acknowledgements

The authors gratefully acknowledge funding provided by the Food Standards Agency (UK) through project E01059: “The applicability of using plasmid DNA as opposed to extracted genomic DNA for quantitative PCR calibration”.

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Authors and Affiliations

  1. Bio-Molecular Innovation, LGC Limited, Queens Road, Teddington, Middlesex, TW11 0LY, UK

    Malcolm Burns, Hernan Valdivia, Paul McDonald & Neil Harris

  2. IRMM, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Retieseweg 111, B-2440, Geel, Belgium

    Philippe Corbisier, Heinz Schimmel & Diana Charels

  3. RHM Technology Ltd., The Lord Rank Centre, Lincoln Road, High Wycombe, Buckinghamshire, HP12 3QR, UK

    Gordon Wiseman, Peter Bowler & Katrina Ohara

  4. Food Standards Agency, Aviation House, 125 Kingsway, London, WC2B 6NH, UK

    Andrew Damant

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  1. Malcolm Burns
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  2. Philippe Corbisier
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  3. Gordon Wiseman
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  4. Hernan Valdivia
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  6. Peter Bowler
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  7. Katrina Ohara
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  8. Heinz Schimmel
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  11. Neil Harris
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Correspondence to Malcolm Burns.

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Burns, M., Corbisier, P., Wiseman, G. et al. Comparison of plasmid and genomic DNA calibrants for the quantification of genetically modified ingredients. Eur Food Res Technol 224, 249–258 (2006). https://doi.org/10.1007/s00217-006-0376-z

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  • DOI: https://doi.org/10.1007/s00217-006-0376-z

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