Rocca Angela, Martin Laurent, Kuuranne Tiia, Ericsson Magnus, Marchand Alexandre, Leuenberger Nicolas
Swiss Laboratory for Doping Analyses, University Center of Legal Medicine, Lausanne and Geneva, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
Analysis Department, Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry, France.
Drug Test Anal. 2022 May;14(5):820-825. doi: 10.1002/dta.3142. Epub 2021 Aug 21.
Continuous erythropoietin receptor activator (CERA) is a third-generation erythropoiesis-stimulating agent that was developed for the treatment of anemia. However, misuse of CERA for doping in endurance sports has been reported. Previous studies have shown blood as the matrix of choice for the detection of CERA, due to its high molecular weight. The use of dried blood spots (DBSs) for anti-doping purposes constitutes a complementary approach to the standard urine and venous blood matrices and could facilitate sample collection and increase the number of blood samples available for analysis due to reduced costs of sample collection and transport. Here, we investigated whether CERA could be indirectly detected in extracts of single DBSs using an erythropoietin-specific immunoassay that is capable of providing results within approximately 2 h. Reconstituted DBS samples were prepared from mixtures of red blood cell pellets and serum samples. The samples were collected in a previous clinical study in which six healthy volunteers were injected with a single, 200 μg dose of CERA. Using a commercially available ELISA kit, CERA was detected in the DBSs with a detection window of up to 20 days post-injection. Furthermore, in order to demonstrate the fitness-for-purpose, three authentic doping control serum samples, which were identified as containing CERA, were analyzed by the presented methodological approach on DBS. The testing procedure described here could be used as a fast and cost-effective method for the detection of CERA abuse in sport.
持续促红细胞生成素受体激活剂（CERA）是一种用于治疗贫血的第三代促红细胞生成剂。然而，已有报道称CERA被滥用于耐力运动中的兴奋剂使用。先前的研究表明，由于CERA分子量较大，血液是检测其的首选基质。使用干血斑（DBS）进行反兴奋剂检测是对标准尿液和静脉血基质的一种补充方法，由于样本采集和运输成本降低，它可以促进样本采集并增加可供分析的血样数量。在此，我们研究了是否可以使用一种能够在约2小时内得出结果的促红细胞生成素特异性免疫测定法，间接检测单个DBS提取物中的CERA。从红细胞沉淀和血清样本的混合物中制备重构的DBS样本。这些样本是在先前的一项临床研究中收集的，该研究中6名健康志愿者被注射了单次200μg剂量的CERA。使用市售的酶联免疫吸附测定试剂盒，在注射后长达20天的检测窗口期内，在DBS中检测到了CERA。此外，为了证明该方法的适用性，对三个经鉴定含有CERA的真实兴奋剂检测对照血清样本，采用本文介绍的DBS方法进行了分析。这里描述的检测程序可以作为一种快速且经济高效的方法，用于检测运动中滥用CERA的情况。
