EPO transgene detection in dried blood spots for antidoping application

EPO transgene detection in dried blood spots for antidoping application / Alexandre Marchand, Ingrid Roulland, Florian Semence, Magnus Ericsson

  • Drug Testing and Analysis 13 (2021) 11-12 (November-December), p. 1888-1896
  • Special Issue: The 39th Manfred Donike workshop on doping analysis
  • PMID: 33961347
  • DOI: 10.1002/dta.3059


The modification of gene expression to treat diseases is a field of research with exponential growth. As doping in sport closely follows emerging therapies, a surveillance of the modification of gene expression to enhance performance is needed. The gene coding for erythropoietin (EPO) is one target of interest. Since 2010, several protocols have been proposed to identify EPO gene doping by focusing on the presence in blood of a transgene that differ in size from the endogenous gene sequence, normally found in the human DNA. In this work, our aim was to validate an easily applicable method for EPO gene doping detection in dried blood spots (DBS). We evaluated the detection of EPO transgene in 20-μl DBS after the spike of a plasmid carrying the EPO transgene in whole blood. Three different DBS were compared: Nucleic-Card™, Whatman® 903, and the volumetric 20-μl VAMS™. Detection was performed with real-time polymerase chain reaction (PCR) and validated with two Taqman assays (one commercial and one custom) specific for the EPO transgene. The initial testing procedure could be done using one assay (custom) and the confirmation using the second one (commercial Taqman) with a final check of the size of the PCR product. Starting from 20-μl dried blood, 1000 copies of EPO transgene could efficiently be detected with the three types of DBS, VAMS showing a slightly better sensitivity. No loss of sensitivity was observed after 1-month storage of DBS at room temperature. This method could be applied to DBS collected during doping controls and allows reanalysis.


Research / Study
7 May 2021
Ericsson, Magnus
Marchand, Alexandre
Roulland, Ingrid
Semence, Florian
Paris, France: Agence Française de Lutte contre le Dopage (AFLD)
Analytical aspects
Blood testing method
DNA analysis
Testing method development
Doping classes
S2. Peptide Hormones, Growth Factors
Erythropoietin (EPO)
Medical terms
Gene Therapy
Document category
Date generated
20 January 2022
Date of last modification
27 January 2022
  • Legal Source
  • Education
  • Science
  • Statistics
  • History
Country & language
  • Country
  • Language
Other filters
  • ADRV
  • Legal Terms
  • Sport/IFs
  • Other organisations
  • Laboratories
  • Analytical aspects
  • Doping classes
  • Substances
  • Medical terms
  • Various
  • Version
  • Document category
  • Document type
Publication period