Development on Confirmation Method for recombinant EPO Analysis in Individuals with c.577del in EPO Gene

Code: 22B06XZ

Upon our previous research, a variant in the EPO gene can encode a variant EPO (VAR-EPO) which has a similar molecule weight with recombinant EPO (rEPO), thus cannot be distinguished using the current method of rEPO analysis. Though WADA has established a detailed procedure for investigation and further analysis, it is a burden for both laboratories, Testing Authoity and WADA, nevertheless, rEPO abused by this variant carriers can be hardly detected. For this situation, we are going to develop an effective confirmation method that can discriminate the wild-type EPO, VAR-EPO and rEPO directly, by removing or eliminating the detection of VAR-EPO using the VAR-specific antibody. Two methods will be developed and validated in this project:: reverse-normal immunopurification and step-recognition western blotting. The rEPO administration study on individuals who carry this varient will also be conducted. The collected positive urine and blood samples will be used to evaluate the method. With out goals, the new method can not only protect the "clean" athlete with that variant but also effectively penalize the rEPO abused athelete whether he or she carries that variant. Another significant advantage, this method would be easily adopted by other WADA accredited anti-doping laboratories.