0000000000046994
AUTHOR
E. Neuberger
Poster Presentations
Administration of recombinant human erythropoietin (rHuEpo) improves performance and hence is subject to abuse by athletes. The detection of rHuEpo doping remains a major challenge at present. The aim of the current study was to investigate whether circulating microRNA (miRNA) can be used for detecting r-HuEpo doping. Twenty trained males received rHuEpo injections of 50 IU∙kg -1 body mass every two days for 4 weeks. Blood was obtained 2 weeks before, during and 4 weeks after administration. For this pilot analysis, plasma miRNA expression was assessed at selected time points using the Affymetrix GeneChip 3.0 and the miScript 384 HC PCR Array (Qiagen). For the Affymetrix microarray data, GC…
Establishment of two quantitative nested qPCR assays targeting the human EPO transgene.
International audience; For ethical and safety reasons it is critical to develop easily implemented assays with high sensitivity and specificity for gene doping surveillance. Two nested quantitative real-time PCR (qPCR) assays were developed that target the human EPO (hEPO) cDNA sequence in a circular form, representative of recombinant adeno-associated viral (rAAV) vector genomes found in vivo. Through an interlaboratory evaluation, the assays were validated and utilized in an in vitro blinded study. These assays are specific and extremely sensitive with a limit of detection (LOD) of 1 copy of circular plasmid DNA and a limit of quantification (LOQ) of 10 to 20 copies in the presence of 50…
Antidoping Science
The ineffectiveness of antidoping programs in elite sport, largely due to human and political factors, is leading to a new resolve and greater transparency of antidoping authorities and those stakeholders interested in drug-free sport. The perception by the public, athletes, and the World Anti-Doping Agency (WADA) of antidoping science and current drug testing programs in elite sport varies widely from "ineffective" to "robust and reliable." Here, we discuss why a careful and considered reevaluation of the underlying premise of antidoping science is needed to bring this unique application of predictive/diagnostic science more in line with other areas of medicine. We show how the validity of…
10th international Luebeck conference on the pathophysiology and pharmacology of erythropoietin and other hemopoietic growth factors
Recombinant erythropoietins (rEPOs) as well as other erythropoiesis-stimulating agents (ESAs) are used for medical treatment of anemia, but likewise ESAs can be misused for performance enhancing purposes in sports. Using small RNAseq, we investigated the effects of low dose administration of rEPO for 6 weeks on the miRNA profile of purified red blood cells (RBCs), in 7 male healthy recreational athletes. Despite lacking most of long RNAs, RBCs contain diverse and abundant miRNAs. For each of the 14 samples 100 ng total RNA, of highly purified RBCs, were used for sequencing with Illumina NextSeq500 system. Following bowtie alignment and mapping of the raw reads, edgeR was used for normalizat…