EPO gene doping test: additional validation with more blood samples and storage protocols

Code: ISF18R01AB 

In this project, we propose to perform additional validation of the EPO gene doping test with blood samples on the conditions for samples storage.  This includes factors affecting centrifugation and thawing, assay performance, improve separation of Tm peaks, storage temperature.  The assay will be validated with the optimal conditions.

The further validation of the EPO gene doping test will facilitate completion of the Technical Document for gene doping detection in blood and implementation of the test in the near future.

Main Findings: 

We performed additional validation of the polymerase chain reaction (PCR)-based erythropoietin (EPO) gene doping test using blood samples. We tested various conditions for prolonged sample storage and determined storage temperatures and duration, as well as sample processing prior to freezing, which do not compromise reliability of detecting the EPO doping gene in blood. We also improved test reliability by using a more suitable form of Uracil-DNA Glycosylase in PCR mix and tested other modifications to the test protocol with the intention to make the test easier and more legally defensible.
The results and outcomes from this project were incorporated in the Test Protocol. We conclude that the PCR-based EPO gene doping test using whole blood samples was fit to be implemented in doping control.