Development, implementation and validation of a test for detecting a dual population of red blood cells.
Objectives
The objective of this study was to develop the capacity to detect a dual population of red blood cells in a blood sample, indicating homologous blood transfusion.
Introduction
Since the 1970s, a large number of investigations have been conducted demonstrating that blood transfusion enhances the performance in high level athletes. The observed improvement is explained by an increase in the mass of hemoglobin resulting in higher oxygenation of muscles. Homologous transfusion consists in injecting the recipient with blood collected from a donor of the same blood group. This method is very risky from a medical point of view, especially if transfusion takes place outside a hospital setting.
Homologous transfusion probably lost its appeal in the 80s when recombinant erythropoietin (EPO) became available. EPO is easier to use, more discrete, less dangerous and just as effective. However, blood transfusions became popular again among certain athletes after year 2000, when an EPO anti-doping test was developed.
A homologous transfusion invariably yields mixed blood with two different red blood cell populations. The two blood types may be fully compatible but differences in blood groups exist even if they have not been detected in tests used prior to the transfusion.
Methods
To reveal the differences in red blood cell populations, immunological labels, specific for certain blood groups were used and the populations were analyzed by flow cytometry. A blind clinical study was carried out to determine, among other parameters, the sensitivity and the specificity of this method. The analysis was conducted on unknown samples containing
0%, 0.5%, 1.5%, 3% or 5% of the second, minor population of red blood cells.
It was also necessary to analyze the red blood cells made available by the industry as part of commercialized serology products .
Results
We have developed and integrated control procedures for specificity and identification capacity of all the antibodies used in this work.
The method displays 100% specificity with respect to the antibodies and the samples, whereas global sensitivity reaches 66.8%. If the samples that contain only 0.5% of the smaller population are ignored, sensitivity reaches 80,5%.
The detection range for this method lasts between 2 and 3 months depending on the antigens associated with the two blood populations.
Publications
- Giraud S, Robinson N, Mangin P, Saugy M. Scientific and forensic standards for homologous blood transfusion anti-doping analyses. Forensic Sci Int. 2008 Jul 18;179(1):23-33
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