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Direct and long-term detection of gene doping in conventional blood samples

Gene Therapy volume 18, pages 225–231 (2011)Cite this article

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Abstract

The misuse of somatic gene therapy for the purpose of enhancing athletic performance is perceived as a coming threat to the world of sports and categorized as ‘gene doping’. This article describes a direct detection approach for gene doping that gives a clear yes-or-no answer based on the presence or absence of transgenic DNA in peripheral blood samples. By exploiting a priming strategy to specifically amplify intronless DNA sequences, we developed PCR protocols allowing the detection of very small amounts of transgenic DNA in genomic DNA samples to screen for six prime candidate genes. Our detection strategy was verified in a mouse model, giving positive signals from minute amounts (20 μl) of blood samples for up to 56 days following intramuscular adeno-associated virus-mediated gene transfer, one of the most likely candidate vector systems to be misused for gene doping. To make our detection strategy amenable for routine testing, we implemented a robust sample preparation and processing protocol that allows cost-efficient analysis of small human blood volumes (200 μl) with high specificity and reproducibility. The practicability and reliability of our detection strategy was validated by a screening approach including 327 blood samples taken from professional and recreational athletes under field conditions.

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Acknowledgements

This project has been carried out with the support of WADA (research grants 06B7PS and 08C17PS). We thank Irina Smirnow and Andrea Schenk (Department of Gastroenterology & Hepatology, Medical Clinic, Eberhard-Karls-University of Tuebingen) for excellent technical assistance.

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Authors and Affiliations

  1. Department of Sports Medicine, Medical Clinic, Eberhard-Karls-University of Tuebingen, Tuebingen, Germany

    T Beiter, A Fragasso, J Hudemann & A M Niess

  2. Department of Gastroenterology & Hepatology, Medical Clinic, Eberhard-Karls-University of Tuebingen, Tuebingen, Germany

    M Zimmermann, M Bitzer & U M Lauer

  3. Department of Sports Medicine, Rehabilitation and Disease Prevention, Johannes Gutenberg-University Mainz, Mainz, Germany

    P Simon

Authors
  1. T Beiter
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  2. M Zimmermann
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  3. A Fragasso
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  4. J Hudemann
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  5. A M Niess
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  6. M Bitzer
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  7. U M Lauer
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  8. P Simon
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Corresponding author

Correspondence to P Simon.

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The authors declare no conflict of interest.

Additional information

The University of Tübingen, Germany has a patent pending for the ‘Detection of transgenic DNA’(PCT/EP2007/003385; http://www.wipo.int/pctdb/en/wo.jsp?WO=2007124861) that relates to the detection of transgenic DNA in a living being and to a kit for performing such a method. A free use without charge of the patent pending procedure has been granted to the World Anti-Doping Agency for the purpose of doping analysis in sports.

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Beiter, T., Zimmermann, M., Fragasso, A. et al. Direct and long-term detection of gene doping in conventional blood samples. Gene Ther 18, 225–231 (2011). https://doi.org/10.1038/gt.2010.122

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  • DOI: https://doi.org/10.1038/gt.2010.122

Keywords

  • gene doping
  • doping detection
  • doping
  • gene transfer
  • sport

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