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PCR-based detection of gene transfer vectors: application to gene doping surveillance

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Abstract

Athletes who illicitly use drugs to enhance their athletic performance are at risk of being banned from sports competitions. Consequently, some athletes may seek new doping methods that they expect to be capable of circumventing detection. With advances in gene transfer vector design and therapeutic gene transfer, and demonstrations of safety and therapeutic benefit in humans, there is an increased probability of the pursuit of gene doping by athletes. In anticipation of the potential for gene doping, assays have been established to directly detect complementary DNA of genes that are top candidates for use in doping, as well as vector control elements. The development of molecular assays that are capable of exposing gene doping in sports can serve as a deterrent and may also identify athletes who have illicitly used gene transfer for performance enhancement. PCR-based methods to detect foreign DNA with high reliability, sensitivity, and specificity include TaqMan real-time PCR, nested PCR, and internal threshold control PCR.

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Acknowledgments

This project was funded by the World Anti-Doping Agency (WADA; R.O.S. and P.M.), United States Anti-Doping Agency (R.O.S.), Partnership for Clean Competition (R.O.S.), and Agence Francaise de Lutte contre le Dopage (P.M.). This work was performed under a Cooperative Agreement between INSERM, AFM, L'Etablissement Francais du Sang, and the University of Florida Center of Excellence for Regenerative Health Biotechnology.

Conflict of interest

R.O.S. is an inventor on patents related to rAAV technology. R.O.S owns equity in a gene therapy company that is commercializing adeno-associated virus for gene therapy applications. To the extent that the work reported in this article increases the value of these commercial holdings, R.O.S. has a conflict of interest.

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Authors and Affiliations

  1. Department of Molecular Genetics and Microbiology, University of Florida, College of Medicine, 1600 SW Archer Road, Gainesville, FL, 32610-0266, USA

    Irene C. Perez, Weiyi Ni, Jennifer Lyles, Philippe Moullier & Richard O. Snyder

  2. Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, 44007, France

    Caroline Le Guiner, Philippe Moullier & Richard O. Snyder

  3. Center of Excellence for Regenerative Health Biotechnology, University of Florida, Alachua, FL, 32615, USA

    Richard O. Snyder

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  1. Irene C. Perez
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  2. Caroline Le Guiner
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  3. Weiyi Ni
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  4. Jennifer Lyles
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  5. Philippe Moullier
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  6. Richard O. Snyder
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Correspondence to Richard O. Snyder.

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Published in the topical collection Anti-doping Analysis with guest editor Christopher Harrison.

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Perez, I.C., Le Guiner, C., Ni, W. et al. PCR-based detection of gene transfer vectors: application to gene doping surveillance. Anal Bioanal Chem 405, 9641–9653 (2013). https://doi.org/10.1007/s00216-013-7264-8

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