Establishing a novel single-copy primer-internal intron-spanning PCR (spiPCR) procedure for the direct detection of gene doping.
So far, the abuse of gene transfer technology in sport, so-called gene doping, is undetectable. However, recent studies in somatic gene therapy indicate that long-term presence of transgenic DNA (tDNA) following various gene transfer protocols can be found in DNA isolated from whole blood using conventional PCR protocols. Application of these protocols for the direct detection of gene doping would require almost complete knowledge about the sequence of the genetic information that has been transferred. Here, we develop and describe the novel single-copy primer-internal intron-spanning PCR (spiPCR) procedure that overcomes this difficulty. Apart from the interesting perspectives that this spiPCR procedure offers in the fight against gene doping, this technology could also be of interest in biodistribution and biosafety studies for gene therapeutic applications.
Beiter
Thomas
T
Department of Sports Medicine, Medical Clinic, University of Tüebingen, Germany.
Zimmermann
Martina
M
Fragasso
Annunziata
A
Armeanu
Sorin
S
Lauer
Ulrich M
UM
Bitzer
Michael
M
Su
Hua
H
Young
William L
WL
Niess
Andreas M
AM
Simon
Perikles
P
D004247:DNA / Q000097:blood; D004300:Doping in Sports / Q000941:ethics / Q000379:methods / Q000517:prevention & control; D004921:Erythropoietin / Q000235:genetics / Q000378:metabolism*; D015316:Genetic Therapy; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000639:trends; D019076:Transgenes / Q000941:ethics / Q000235:genetics*; D042643:Vascular Endothelial Growth Factor D / Q000235:genetics / Q000378:metabolism*
D042643:Vascular Endothelial Growth Factor D; D004921:Erythropoietin; D004247:DNA