GASEPO2- A software tool for analysis of EPO images after isoelectric focusing and double blotting
Since the development of an analytical method for the direct detection of the application of recombinant erythropoietin in urine samples by Lasne et. al. (Nature, 2000, 405, 635), great efforts have been made to implement the method in different laboratories and to standardize and harmonize the interpretation of the data obtained by isoelectric focussing, double blotting and chemiluminiscence detection. Up to now no final decision has been made by the labs accredited for the cited method, which sets of criteria for positivity are to be applied for the interpretation of the gels. The use of different software products with different tools of data interpretation interferes the search for a common basis of gel interpretation. The aim of this project proposal is to develop a reliable and easy-to-use software package called GASEPO2, specially designed for quantification of recombinant erythropoietin in urine samples as a standardized and unified tool for the use across doping-control laboratories worldwide thus providing for international harmonization of EPO analysis. This software is an advancement of the precursor software GASEPO1, designed as research prototype in 2003. The development includes research in design of decision criteria based on image information as well as in mathematical methods of image analysis including image segmentation and 3D visualisation. By using the concept of band segmentation for the differentiation between band and background, the isolated single band serves as basis for further calculation rather than a 1 D transformation of the entire lane. GASEPO2 will equally well handle images taken by a dedicated digital camera for chemiluminescence or scanned from a photographic film. The software will come with a validation package that allows auditing. The implementation of alternative evaluation methods in the software will contribute to and facilitate the process of finding and harmonizing the decision criteria.
The Project fulfilled the original goals, namely: • A software tool has been created that has been generally accepted by the doping control community and is now being routinely used in 75% of all laboratories doing epo-analytics. • Thus, the product of the Project contributed directly to the superordinate aim of standardization and harmonization of the epo analytics. • An effective epo-analytics expert network has been created. • Scientific know-how has been extended and disseminated via a number of scientific publications. • WADA-funded research in doping control has been promoted by a number of project-related presentations for broader public.