PCR-based detection of gene transfer vectors: application to gene doping surveillance
- PDF / 374,774 Bytes
- 13 Pages / 595.276 x 790.866 pts Page_size
- 13 Downloads / 159 Views
REVIEW
PCR-based detection of gene transfer vectors: application to gene doping surveillance Irene C. Perez & Caroline Le Guiner & Weiyi Ni & Jennifer Lyles & Philippe Moullier & Richard O. Snyder
Received: 20 May 2013 / Revised: 3 July 2013 / Accepted: 16 July 2013 / Published online: 4 August 2013 # Springer-Verlag Berlin Heidelberg 2013
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.
Keywords Nested PCR . Real-time PCR . Erythropoietin . Gene transfer . Internal threshold control PCR . Vector
Published in the topical collection Anti-doping Analysis with guest editor Christopher Harrison. I. C. Perez : W. Ni : J. Lyles : P. Moullier : R. O. Snyder Department of Molecular Genetics and Microbiology, University of Florida, College of Medicine, 1600 SWArcher Road, Gainesville, FL 32610-0266, USA C. Le Guiner : P. Moullier : R. O. Snyder Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes 44007, France R. O. Snyder (*) Center of Excellence for Regenerative Health Biotechnology, University of Florida, Alachua, FL 32615, USA e-mail: [email protected]
Introduction Advances in recombinant DNA and protein technology as well as in gene transfer have led to new treatments for diseases. Unfortunately, new drugs developed from these advancements such as recombinant human growth hormone and erythropoietin (EPO) have been abused by athletes. In recent years, improvements in assays to detect doping have identified more athletes who dope, some of whose previous samples tested negative, and high-profile admissions confirm widespread doping in cycling and track and field sports [1, 2]. Gene doping in sports was banned in 2003 by the World AntiDoping Agency (WADA), and the 2013 WADA Prohibited List defines gene doping as “the transfer of polymers of nucleic acids or nucleic acid analogues” and/or “the use of normal or genetically modified cells” with the potential to enhance sport performance [3]. With the advancements made in gene transfer for the therapeutic treatment of different genetic disorders, there ha
Data Loading...
