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Detection of EPO injections using a rapid lateral flow isoform test

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Abstract

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.

The lateral flow isoform test, in a dipstick format, can distinguish different types of EPO (blue balls) due to the interaction of their glycosylated structures with the lectin wheat germ agglutinin (WGA), which is the 1st zone to pass. EPO that have succeeded to pass the WGA zone will be captured by EPO-specific antibodies found in the subsequent zone. The percentage of passed EPO is calculated by also measuring the total amount of EPO in the sample using a dipstick with inactive WGA zone.

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Acknowledgments

The authors thank Maria Andrén, Niklas Rollborn, Malin Drevin, Mikael Lönnberg and Trikien Quach for technical assistance. The authors are grateful for the financial support for the development of the MAIIA test from VINNOVA (The Swedish Governmental Agency for Innovation Systems) in the project P25917.

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Authors and Affiliations

  1. Department of Chemistry-BMC, Analytical chemistry, Uppsala University, 751 24, Uppsala, Sweden

    M. Lönnberg

  2. Center for Integrative Human Physiology (ZIHP), Institute of Physiology, University of Zurich, 8006, Zürich, Switzerland

    C. Lundby

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  1. M. Lönnberg
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  2. C. Lundby
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Correspondence to M. Lönnberg.

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Published in the topical collection Anti-doping Analysis with guest editor Christopher Harrison.

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Lönnberg, M., Lundby, C. Detection of EPO injections using a rapid lateral flow isoform test. Anal Bioanal Chem 405, 9685–9691 (2013). https://doi.org/10.1007/s00216-013-6997-8

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  • DOI: https://doi.org/10.1007/s00216-013-6997-8

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