Subscribe to RSS
DOI: 10.1055/s-2007-965076
© Georg Thieme Verlag KG Stuttgart · New York
Detection of Homologous Blood Transfusion
Publication History
accepted after revision January 24, 2007
Publication Date:
05 July 2007 (online)
![](https://www.thieme-connect.de/media/sportsmed/200708/lookinside/thumbnails/10.1055-s-2007-965076-1.jpg)
Abstract
The aim of the present study was to improve and validate a flow cytometric method for the detection of homologous blood transfusion in doping control analysis. A panel of eight different primary antibodies and two different phycoerythrin-conjugated secondary antibodies was used for the detection of different blood populations. The flow cytometer used in this study was the BD FACSArray® instrument. Mixed red blood cell populations were prepared from phenotype known donors. Linearity, specificity, recovery, precision, robustness and interday-precision were tested for every primary antibody used in the presented assay. The technique of signal amplification was utilized for an improved separation of antigens with weak or heterozygous expression to improve the interpretation of histograms. The resulting method allowed to clearly identify mixed red blood cell populations in homologous blood transfusion samples containing 0.3 - 2.0 % of donor blood.
Key words
doping - sport - blood transfusion - flow cytometry
References
- 1 Ashenden M. A strategy to deter blood doping in sport. Haematologica. 2002; 87 225-234
- 2 Cazzola M. A global strategy for prevention and detection of blood doping with erythropoietin and related drugs. Haematologica. 2000; 85 561-653
- 3 Ekblom B, Berglund B. Effect of erythropoietin administration on maximal aerobic power. Scand J Mec Sci Sports. 1991; 1 88-93
- 4 Ekblom B. Blood doping and erythropoietin. Am J Sports Med. 1996; 24 40-42
- 5 Gore C J, Parisotto R, Ashenden M J, Stray-Gundersen J, Sharpe K, Hopkins W, Emslie K R, Howe C, Trout G J, Kazlauskas R, Hahn A G. Second-generation blood tests to detect erythropoietin abuse by athletes. Haematologica. 2003; 88 333-344
- 6 Gottwald W. Statistik für Anwender. Weinheim; Wiley-VCH 2004: 111-112
- 7 Griffin G D, Lippert L, Dow N S, Berger T A, Hickman M R, Salata K F. A flow cytometric method for phenotyping recipient red cells following transfusion. Transfusion. 1994; 34 233-237
- 8 Heinicke K, Heinicke I, Schmidt W, Wolfarth B. A three-week traditional altitude training increases haemoglobin mass and red cell volume in elite biathlon athletes. Int J Sports Med. 2005; 26 350-355
- 9 Heinicke K, Wolfarth B, Winchenbach P, Biermann B, Schmid A, Huber G, Friedmann B, Schmidt W. Blood volume and haemoglobin mass in elite athletes of different disciplines. Int J Sports Med. 2001; 22 504-512
- 10 Lasne F, de Ceaurritz J. Recombinant erythropoietin in urine. Nature. 2000; 405 635
- 11 Leigh-Smith S. Blood boosting. Br J Sports Med. 2004; 38 99-101
- 12 Lippi G, Banfi G. Blood transfusions in athletes. Old dogmas, new tricks. Clin Chem Lab Med. 2006; 44 1395-1402
- 13 Malcovati L, Pascutto C, Cazzola M. Hematologic passport for athletes competing in endurance sports: a feasibility study. Haematologica. 2003; 88 570-581
- 14 Nelson M, Ashenden M, Langshaw M, Popp H. Detection of homologous blood transfusion by flow cytometry: a deterrent against blood doping. Haematologica. 2002; 87 881-882
- 15 Nelson M, Cooper S, Nakhla S, Smith S, King M, Ashenden M. Validation of a test designed to detect blood-doping of elite athletes by homologous transfusion. Aust J Med Sci. 2004; 25 27-33
- 16 Nelson M, Popp H, Ashenden M. Proof of homologous blood transfusion through quantification of blood group antigens. Haematologica. 2003; 88 1284-1295
- 17 Nelson M. An overview of the use of flow cytometry in the analysis of mixed red cell populations. Pathology. 1999; 31 191-198
- 18 Nelson M. Analysis of heterogenous red cell populations by flow cytometry. Methods Cell Sci. 2002; 24 19-25
- 19 Sharpe K, Ashenden M, Schumacher Y O. A third generation approach to detect erythropoietin abuse in athletes. Haematologica. 2006; 91 356-363
- 20 Thevis M, Ogorzalek Loo R, Loo J A, Schänzer W. Doping control analysis of bovine hemoglobin-based oxygen therapeutics in human plasma by LC-electrospray ionization-MS/MS. Anal Chem. 2003; 75 3287-3293
- 21 Varlet-Marie E, Ashenen M, Lasne F, Sicart M T, Marion B, de Ceaurritz J, Audran M. Detection of hemoglobin based oxygen carriers in human serum for doping analysis: confirmation by size exclusion HPLC. Clin Chem. 2004; 50 723-731
- 22 WADA, The World Anti Doping Code .The 2006 prohibited list, international standard. http://www.wada-ama.org/rtecontent/document/Prohibited_List_2006_EN_FR_SP.pdf. Access date: 18.08.2006
Prof. Mario Thevis
Center for Preventive Doping Research
Institute of Biochemistry
German Sport University Cologne
Carl-Diem-Weg 6
50933 Cologne
Germany
Fax: + 49 22 14 97 32 36
Email: thevis@dshs-koeln.de