Development of an assay for genotyping of blood groups by polymerase chain reaction
Cardiff Metropolitan University
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The use of DNA based tests in the prediction of blood group phenotype has been made possible since the identification and sequencing of the genes which encode 29 of the 30 blood group systems. Classical methods of determining phenotype are still widely used and in the majority of cases appropriate. In some patient groups and for some blood group systems serological phenotyping can be problematic. In these instances the limitations of serological methods may be overcome by the use of DNA based assays to predict phenotype. The aim of the study was to develop a polymerase chain reaction (PCR) based assay that would allow the accurate prediction of Rh phenotype using sequence specific primers to detect regions of the RHD and RHCE genes in a single assay. The study was successful in developing an in house platform for genotyping by PCR. The sensitivity and specificity of the primers was established by testing a panel of samples of known serological phenotype. 50 samples were tested and interpreted without prior knowledge of serological phenotype. Genotype results were concordant with phenotype in 100% of the test samples. During validation two Rh D negative, Rh C positive samples produced unexpected genotyping results. Research of current scientific literature provided probable explanations for these results.
MSc Biomedical Sciences (Immunohaematology)
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