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Gene doping detection: evaluation of approach for direct detection of gene transfer using erythropoietin as a model system

Gene Therapy volume 17, pages 1022–1032 (2010)Cite this article

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Abstract

As clinical gene therapy has progressed toward realizing its potential, concern over misuse of the technology to enhance performance in athletes is growing. Although ‘gene doping’ is banned by the World Anti-Doping Agency, its detection remains a major challenge. In this study, we developed a methodology for direct detection of the transferred genetic material and evaluated its feasibility for gene doping detection in blood samples from athletes. Using erythropoietin (EPO) as a model gene and a simple in vitro system, we developed real-time PCR assays that target sequences within the transgene complementary DNA corresponding to exon/exon junctions. As these junctions are absent in the endogenous gene due to their interruption by introns, the approach allows detection of trace amounts of a transgene in a large background of the endogenous gene. Two developed assays and one commercial gene expression assay for EPO were validated. On the basis of ability of these assays to selectively amplify transgenic DNA and analysis of literature on testing of gene transfer in preclinical and clinical gene therapy, it is concluded that the developed approach would potentially be suitable to detect gene doping through gene transfer by analysis of small volumes of blood using regular out-of-competition testing.

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Acknowledgements

This work was supported by the Australian Government through the Anti-Doping Research Program (ADRP) of the Department of Communications, Information Technology and the Arts to AB and KRE. The work undertaken by the authors has been done pursuant to the strict provisions of the Crown Use exemption under the Patents Act 1990 (Cth). It is the responsibility of the reader to acknowledge and respect the rights or claims of third parties. We thank Dr Ian E. Alexander from Gene Therapy Research Unit, The Children's Hospital Westmead and Children's Medical Research Institute, Westmead, New South Wales, Australia for support and fruitful discussions, Dr Fabrice Magnino from Integrated Sciences Pty Ltd for help with design and optimisation of assays; and Dr Kate Griffiths and Dr Ray Kazlauskas, National Measurement Institute, for critically reading the manuscript and providing constructive advice.

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  1. Bioanalysis, National Measurement Institute, New South Wales, Australia

    A Baoutina, T Coldham, G S Bains & K R Emslie

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  1. A Baoutina
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  2. T Coldham
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  3. G S Bains
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Correspondence to A Baoutina.

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Baoutina, A., Coldham, T., Bains, G. et al. Gene doping detection: evaluation of approach for direct detection of gene transfer using erythropoietin as a model system. Gene Ther 17, 1022–1032 (2010). https://doi.org/10.1038/gt.2010.49

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