The use of molecular typing to improve the quality of reagent red blood cells
Cardiff Metropolitan University
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Reagent red blood cells (RBCs) selected for antibody screening and identification red cell panels are comprehensively phenotyped for all significant blood group antigens. The classical method of testing for blood group antigens and antibodies is haemagglutination. With the exception of RhD, Duffy (Fy) and S antigens haemagglutination-based determination of RBC phenotype provides a reliable method of determining antigen dosage. Polyclonal antibodies, produced in response to transfusion or pregnancy, vary widely in avidity and strength; some react strongly with reagent RBCs carrying the appropriate antigen, whereas others show a 'dosage' effect and will react only with RBCs with a homozygous expression of the antigen. If presumed homozygous RBCs are used this may compromise the ability to detect weak antibodies due to their reduced antigen expression. The aim of this study was to develop a polymerase chain reaction (PCR) based assay to determine the FY genotype with a view to improving the quality of red cell panels used for antibody identification by ensuring accurate determination of homozygous expression of the antigens. The specificity of the primers used was validated by testing a panel of samples of known serological phenotype. Two of the 75 DNA samples genotyped exhibited weakening alleles and on one occasion the serological phenotype did not agree with the resulting genotype.
MSc Biomedical Science
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