Blood online
Home About Blood Authors Subscriptions Permission Advertising Public Access contact us
 

 
Advanced
Current Issue
First Edition
Future Articles
Archives
Submit to Blood
Search
American Society of Hematology
Meeting Abstracts
Email Alerts
Blood, 1 September 2006, Vol. 108, No. 5, pp. 1778.

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Right arrow Rights and Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Catlin, D.
Right arrow Articles by Starcevic, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Catlin, D.
Right arrow Articles by Starcevic, B.
Related Collections
Right arrowRelated Article in Blood Online
Social Bookmarking
 Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

arrow to previous article Previous Article  |  Table of Contents  |  Next Article next article arrow

CORRESPONDENCE

To the editor:

False-positive Epo test concerns unfounded

The brief report by Beullens et al1 is misleading regarding the urine test that the World Anti-Doping Agency (WADA) uses to detect recombinant human erythropoietin (rhEpo). The WADA-recommended test is based on immunoelectorphoresis and double blotting (IEF/DB), and was developed by Lasne and de Ceaurriz in 2000.2

Our WADA-accredited laboratory has performed the IEF/DB test for rhEpo on more than 6800 urine samples, including more than 2600 doping control samples from athletes. Of the latter, we have reported 9 positive cases for rhEpo: 3 of these have publicly confessed to using rhEpo, 3 have accepted penalties, the physician of a seventh has been indicted for distribution of rhEpo, and 2 maintained their innocence but lost on appeal.

We take issue with Beullens's use of the term "false positive" because, as the authors emphasize, the compound they are discussing is not rhEpo. If the compound detected can be identified as not rhEpo, then it cannot cause a false positive. This term sensationalizes an otherwise interesting case report that could in due course contribute to the body of science.

The criteria used by the WADA laboratories are well known and readily available.3 Beullens et al do not state the criteria they used to make the "false-positive" claim. Using the WADA criteria, the "false-positive" electropherogram1(Fig1A) is clearly negative. Moreover, they do not include a negative or a positive urine quality-control sample. Forensic testing results are normally accompanied by a comprehensive documentation package that supports the conclusion. A report such as this that raises a profound issue (false accusations against an athlete) at a minimum requires far more documentation.

Another important but unexplained issue is the nature of the compound that appears to migrate in the same general region as rhEpo and is characterized by bands. The pH range of the ampholytes is needed in order to fully interpret the data. In the left panel of their Figure 1A, the epoietin-beta lane shows 3 faint bands and possibly a very weak fourth band. The bands in the darbepoetin lane are overly dense. Knowing the pH range of the ampholytes might explain why the darbepoetin region seems closer to the rHuEPO region than we customarily observe (compare with Figure 1 here).

The right panel of their Figure 1A shows an apparent protein with bands, but it does not look like a typical rhEpo positive (Figure 1 here). Further, 1 and maybe 2 of the bands migrate more basically than the most basic epoietin-beta band. Under the WADA rules, the identification criteria are not met.


Figure 1
View larger version (75K):
[in this window]
[in a new window]
 
Figure 1.. Electropherogram showing a darbepoetin alfa/rhEpo standard (left lane) and human quality control urines. The darbepoetin alfa/rhEpo standard is on the left, the negative human quality control urine (from a known rhEpo-free donor) is in the middle, and the positive human quality control urine (pooled from known donors on rhEpo) is on the right.

 
Finally, the athlete has a puzzling renal disease characterized by a concentrating defect and an excessive number of casts that apparently does not interfere with his athletic prowess. He should have a full nephrology evaluation.

Don Catlin, Gary Green, Michael Sekera, Paul Scott, and Borislav Starcevic

Correspondence: Don Catlin, Department of Molecular and Medical Pharmacology, UCLA Olympic Analytical Laboratory, University of California, Los Angeles, Los Angeles, CA 90025; e-mail: dcatlin{at}ucla.edu.

References

  1. Beullens M, Delanghe JR, Bollen M. False-positive detection of recombinant human erythropoietin in urine following strenuous physical exercise. Blood. 2006;107: 4711-4713.[Abstract/Free Full Text]

  2. Lasne F, de Ceaurriz J. Recombinant erythropoietin in urine. Nature. 2000;405: 635.[Medline] [Order article via Infotrieve]

  3. Catlin D, Nissen-Lie G, Howe C, et al. Harmonization of the method for the identification of epoetin alfa and beta (EPO) and darbepoetin alfa (NESP) by IEF, double blotting, and chemiluminescent detection. WADA technical document TD2004EPO. October 15, 2004. http://www.wada-ama.org/rtecontent/document/td2004epo_en.pdf. Accessed July 6, 2006.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?

Related Article in Blood Online:

False-positive detection of recombinant human erythropoietin in urine following strenuous physical exercise
Monique Beullens, Joris R. Delanghe, and Mathieu Bollen
Blood 2006 107: 4711-4713. [Abstract] [Full Text] [PDF]



This article has been cited by other articles:


Home page
J. Appl. Physiol.Home page
C. Lundby, N. J. Achman-Andersen, J. J. Thomsen, A. M. Norgaard, and P. Robach
Testing for recombinant human erythropoietin in urine: problems associated with current anti-doping testing
J Appl Physiol, August 1, 2008; 105(2): 417 - 419.
[Abstract] [Full Text] [PDF]


Home page
J. Appl. Physiol.Home page
J. R. Delanghe and M. J. Joyner
Testing for recombinant human erythropoietin
J Appl Physiol, August 1, 2008; 105(2): 395 - 396.
[Full Text] [PDF]


This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Right arrow Rights and Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Catlin, D.
Right arrow Articles by Starcevic, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Catlin, D.
Right arrow Articles by Starcevic, B.
Related Collections
Right arrowRelated Article in Blood Online
Social Bookmarking
 Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

 click for free articles
home about blood authors subscriptions permissions advertising public access contact us
  Copyright © 2006 by American Society of Hematology         Online ISSN: 1528-0020