Detection of microdoses of rhEPO with the MAIIA test

Abstract

The detection of recombinant human erythropoietin (rhEPO) is difficult and becomes more challenging when only microdoses are administered intravenously. Twenty-three subjects were divided into two groups: EPO group (n = 7) and CONTROL group (n = 16). Seven urine and blood samples per subject were collected at least 5 days apart to determine within- and between-subject standard deviations in the percentage of migrating isoforms by the MAIIA test. Six injections of 50 IU/kg bw (boosting dosage) of epoetin beta (Neorecormon, Roche Diagnostics, Hvidovre, Denmark) were performed intravenously during a 3-week period, followed by two microinjections of only 10 IU/kg bw. Blood and urine samples were collected 2, 6, 12, and 72 h after the microinjection, as well as 72 h after the last boosting dose. Sensitivities and specificities of the MAIIA test were examined by absolute and passport thresholds. Sensitivity was 100% for at least 12 h after the microinjection, with ∼30% of plasma samples still exceeding the 99.9% passport threshold 72 h after a microinjection. The specificity was higher for the passport approach compared to the absolute approach, but there were no differences in sensitivities between approaches or between specimens (urine and plasma). We conclude that the MAIIA test shows potential for detecting very small doses of rhEPO.

New insights for identification of doping with recombinant human erythropoietin micro‐doses after high hydration

“…[8,9] We chose a slightly higher but reasonably small 900 IU dose as well as a 500 IU dose of Eprex (11-6.5 IU/kg) for our study. Two situations were tested in this study: one when a micro-dose was injected after an initial step of repeated injections with normal doses of rhEPO and one when just a single micro-dose was injected.…”

Blood manipulation: current challenges from an anti-doping perspective

“…In a recent study, subjects were administered 50 IU/kg of epoetin beta IV twice weekly for 3 weeks, followed by 2 microdosage injections of 10 IU/kg body weight. 29 This administration scheme increased the Hb from 0.6 to 1.8 g/dL on the individual level. When the target Hb has been reached, administration is titrated through microdosing to maintain the supraphysiological Hb and a physiologically normal amount of reticulocytes in the circulation.…”

An integrative ‘Omics’ solution to the detection of recombinant human erythropoietin and blood doping

“…Although the ABP approach can identify abnormal enhanced erythropoiesis regardless of the method used, such as autologous blood transfusion, as shown by Pottgiesser et al 29 in a field-like longitudinal blinded setting, the sensitivity of the ABP has been questioned 30. In order to minimise the risk of being caught via the ABP, it is well known that some athletes are now using ‘microdoses’ of rHumanEPO which allegedly range from 10 to 40 IU/kg body mass 16 20 31. Microdoses of rHumanEPO aim (1) to increase haemoglobin mass (Hb mass ) while avoiding large fluctuations in the ABP blood markers as well as minimising the detection window for conventional direct methods and/or (2) to ‘normalise’ the ABP blood markers after blood manipulations such as autologous blood transfusion 20 31.…”

“…In order to minimise the risk of being caught via the ABP, it is well known that some athletes are now using ‘microdoses’ of rHumanEPO which allegedly range from 10 to 40 IU/kg body mass 16 20 31. Microdoses of rHumanEPO aim (1) to increase haemoglobin mass (Hb mass ) while avoiding large fluctuations in the ABP blood markers as well as minimising the detection window for conventional direct methods and/or (2) to ‘normalise’ the ABP blood markers after blood manipulations such as autologous blood transfusion 20 31. In addition, variation in ABP haematological parameters due to factors such as training or hypoxia exposure can influence the interpretation of the ABP results 32.…”