Extended blood grouping of blood donors with automatable PCR-ELISA genotyping.
BACKGROUND
In the past 10 years, PCR-based methods have been described to allow the detection of gene polymorphisms responsible for many blood group antigens. These methods are routinely used to test samples of fetal origin and to resolve serologic discrepancies. Another interesting application of blood group genotyping could be the extended typing of blood donors for minor antigens to facilitate the procurement of compatible blood for alloimmunized patients.


METHODS
PCR-based tests have been modified to allow multiplex amplification of specific fragments of blood group genes and the convenient detection of hybridized amplicons by ELISA in a microplate format.


RESULTS
The results obtained show that fragments of the Rh (D, c, C, e, E), Kell (K, k), Duffy (Fya, Fyb), and Kidd (Jka, Jkb) genes could be amplified along with controls in multiplex PCR reactions. Labeling of amplicons with digoxigenin allowed their solid-phase detection in microplate wells previously coated with individual blood group-specific oligonucleotides. A comparative study performed with 100 individuals showed a 99.7 percent concordance between genotypes and phenotypes for the 11 antigens assayed, with only three discrepant Fyb genotypes.


CONCLUSIONS
Extended genotyping could be performed once on regular donors and confirmed when needed by standard serologic RBC assays. The format of these tests will allow easy automation of the procedure including the interpretation and downloading of the results with existing ELISA software.
St-Louis
Maryse
M
Department of Research and Development, Héma-Québec, Sainte-Foy, Quebec, Canada. mstlouis@hema-quebec.qc.ca
Perreault
Josée
J
Lemieux
Réal
R
D001331:Automation; D001782:Blood Donors; D001789:Blood Group Antigens; D004375:Duffy Blood-Group System / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005784:Gene Amplification; D005838:Genotype; D006801:Humans; D007519:Isoantigens / Q000032:analysis; D007626:Kell Blood-Group System / Q000276:immunology; D007667:Kidd Blood-Group System / Q000276:immunology; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012204:Rh-Hr Blood-Group System / Q000276:immunology
D001789:Blood Group Antigens; D004375:Duffy Blood-Group System; D007519:Isoantigens; D007626:Kell Blood-Group System; D007667:Kidd Blood-Group System; D012204:Rh-Hr Blood-Group System