To extend PCR-based gene doping detection to more targets

Code: 21E04AB

This year WADA published the first version of Guidelines for Gene Doping Detection. The Guidelines apply to testing for doping genes using polymerase chain reaction (PCR). A PCR-based test developed in our laboratory and approved by WADA to be included in doping control has high sensitivity and is easy to use. Yet, as technologies for genetic measurements continue to develop, anti-doping scientists are employing them to develop alternative methods for gene doping testing.

Recently, we developed a novel method for gene doping detection based on massively parallel sequencing (MPS). This technology is rapidly expanding into clinical care with potential applications in diagnostics and therapeutics. The conceivable strength of the MPS methodology in gene doping testing is that it provides sequence-based information, which may be more difficult to challenge legally.

The current project aims to further validate the developed MPS method that simultaneously targets five genes, likely candidates for doping, but can be adapted for other doping genetic material. We will validate the full experimental protocol, the standard material that was designed and produced for quality controls, and the sophisticated bioinformatics pipeline that was developed for the test. We will also optimise DNA extraction from blood to improve recovery and purity of doping genes to increase reliability of the test. By analysing identical samples using the MPS- and PCR-based methods, we will compare the two tests for detecting five doping genes in terms of sensitivity, cost, ease and reliability.

We expect that on completion of the project and using our experience in developing a gene doping test capable to meet WADA’s accreditation requirements, a prototype test for gene doping detection based on MPS technology will be developed. Project’s outcomes will assist WADA in formulating a plan to transfer the method to WADA-accredited laboratories for implementation in routine anti-doping testing.