Evaluation of the MAIIA dipstick test to detect recombinant human erythropoietin in plasma

Ashenden, Michael; Sharpe, Ken; Garnham, Andrew; Gore, Christopher J.

doi:10.1016/j.jpba.2012.04.025

Cited by 11 publications

(14 citation statements)

References 9 publications

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“…Compared to the recent study by Ashenden et al. (), the BS and WS SDs in our study were somewhat lower than in the previous study (Ashenden et al., ). A lower BS SD indicates that our group of subjects was more homogenous, but that a more heterogeneous group with subjects of, e.g., different training status, gender, fitness level, age, etc., might produce a greater BS SD.…”

Section: Discussioncontrasting

confidence: 97%

“…In a recent study by Ashenden et al. (), the sensitivity of a “Swedish dip‐stick test”, the so‐called MAIIA EPO Test , for detecting small amounts of rhEPO, was examined. To replicate the modus operandi of cheating athletes, small to moderate doses (10–40 IU/kg bw) of epoetin beta were injected intravenously and blood samples obtained at different time points.…”

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confidence: 99%

“…The thresholds in the study by Ashenden et al. () were based on a very limited number of control samples and the sensitivities therefore associated with some degree of uncertainty. From a statistical point of view, a larger number of control samples will increase the precision of the estimated SDs and hence the robustness of the estimated thresholds.…”

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confidence: 99%

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Detection of microdoses of rhEPO with the MAIIA test

Mørkeberg

Sharpe

Karstoft

et al. 2013

Scandinavian Med Sci Sports

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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.

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Section: Discussioncontrasting

confidence: 97%

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confidence: 99%

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Detection of microdoses of rhEPO with the MAIIA test

Mørkeberg

Sharpe

Karstoft

et al. 2013

Scandinavian Med Sci Sports

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“…[20] We wanted to test the sensitivity of the MAIIA isoform assay on samples of both blood and urine collected in the first few days following intravenous administration of microdoses of rhEPO. [20] We wanted to test the sensitivity of the MAIIA isoform assay on samples of both blood and urine collected in the first few days following intravenous administration of microdoses of rhEPO.…”

Section: Introductionmentioning

confidence: 99%

Detection of recombinant EPO in blood and urine samples with EPO WGA MAIIA, IEF and SAR‐PAGE after microdose injections

Dehnes

Shalina

Myrvold

2013

Drug Testing and Analysis

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The misuse of microdoses of performance enhancing drugs like erythropoietin (EPO) constitutes a major challenge in doping analysis. When injected intravenously, the half-life of recombinant human EPO (rhEPO) like epoetin alfa, beta, and zeta is only a few hours and hence, the window for direct detection of rhEPO in urine is small. In order to investigate the detection window for rhEPO directly in blood and urine with a combined affinity chromatography and lateral flow immunoassay (EPO WGA MAIIA), we recruited nine healthy people who each received six intravenously injected microdoses (7.5 IU/kg) of NeoRecormon (epoetin beta) over a period of three weeks. Blood and urine samples were collected in the days following the injections and analyzed with EPO WGA MAIIA as well as the current validated methods for rhEPO; isoelectric focusing (IEF) and sarcosyl polyacrylamide gel electrophoresis (SAR-PAGE). For samples collected 18 h after a microdose, the sensitivity of the EPO WGA MAIIA assay was 100% in plasma and 87.5% in urine samples at the respective 98% specificity threshold levels. In comparison, the sensitivity in plasma and urine was 75% and 100%, respectively, with IEF, and 87.5% in plasma and 100% in urine when analyzed with SAR-PAGE. We conclude that EPO WGA MAIIA is a sensitive assay for the detection of rhEPO, with the potential of being a fast, supplemental screening assay for use in doping analysis.

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“…The isoform methods rely on that large numbers of samples from a reference population have been tested to select the most suitable cut-off limits for achieving highest possible specificity and sensitivity for detecting a potential doping misuse with rhEPO. With the One of the recently presented developed EPO WGA MAIIA test kit [11,15,16]EPO isoform methods show quantitative values for EPO isoforms are obtained, the EPO WGA MAIIA test kit [11,15,16], by using percentage of migrated isoforms (PMI) as unit. Quantification allows the use of confidence limits (CL) calculated from the results for the reference population, and makes between method comparison possiblepossible.…”

Section: Introductionmentioning

confidence: 99%

Detection of EPO injections using a rapid lateral flow isoform test

Lönnberg

Lundby

2013

Anal Bioanal Chem

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Misuse of recombinant human erythropoietin (rhEPO) is a major concern in competitive sports, and the implementation of tests allowing for higher detection rates than what current tests are capable of is required. In this study, a novel lateral flow EPO isoform test kit, EPO WGA MAIIA, is evaluated on the basis of plasma and urine samples obtained from eight healthy males in connection with a 28-day rhEPO injection period. rhEPO was injected every other day during the first 14 days of the study, and the method proved to be 100 % effective in detecting rhEPO in the concomitantly obtained samples. Seven days after the last injection, three positive (>99.99 % confidence limit (CL)) subjects were found. When using 99 % CL as the cut-off limit, six of the eight subjects (75 %) were found to be suspected of doping. Samples obtained 14 and 21 days after the last injection showed no detectable trace of rhEPO. A previous study using indirect methods to determine EPO doping on the same samples indicated only that two of the subjects had suspicious values 7-21 days after the last injection. We propose implementing the easy to-use EPO WGA MAIIA test as an initial screening procedure in anti-doping work to (1) increase the detection rate of potential rhEPO doping athletes and (2) allow for a 10-to 20-fold higher analytical rate than what is possible today. days rhEPO was injected every second day during the first 14 days of the study and the method proved to be very effective (xx out of xx) in detecting thisdoping infor the concomitantly obtained samples collected prior to injection. Samples obtained 14 and 21 days after the last injection showed no detectable rhEPO. Seven days after the last injection three positive (>99.99% CL) subjects were found with the test kit. When using 99% CL as cut-off limit, six of the eight subjects (75%) were suspectedfound to be suspicious for doping.Samples obtained 14 and 21 days after the last injection showed no detectable trace of rhEPO.A previous study using indirect methods to determinefor detecting EPO doping on the same samples indicated only that two of the subjects hadwere suspicious valuespositive during 7-21 days after the last injection.We proposesuggest implementing the easy to-use EPO WGA MAIIA rapid lateral flow test as an initial screening procedure in anti-doping work to 1) increase the detection rate of potential rhEPO doping athletes and 2) allow for analysis of a ten to twenty folds higher analytical rate than what is possible todaymore samples than what is possible today.

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Evaluation of the MAIIA dipstick test to detect recombinant human erythropoietin in plasma

Cited by 11 publications

References 9 publications

Detection of microdoses of rhEPO with the MAIIA test

Detection of microdoses of rhEPO with the MAIIA test

Detection of recombinant EPO in blood and urine samples with EPO WGA MAIIA, IEF and SAR‐PAGE after microdose injections

Detection of EPO injections using a rapid lateral flow isoform test

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