An investigation into blood group genotype and phenotype: An evidence base for future strategies for donor and patient management

Abstract

Exposure to foreign red blood cell (RBC) antigens can result in antibody production and clinically significant adverse consequences, namely in transfusion, haemolytic transfusion reactions, or in pregnancy, haemolytic disease of the foetus and newborn. This thesis investigated genotyping and/or phenotyping of RBC antigens with the aim of improving the safety of transfusion and pregnancy. The overarching hypothesis of the studies was that the application of non-traditional methods to investigate RBC antigen status would lead to more sensitive characterisation of RBCs. The major outcomes of the studies within this thesis included identification of a nonconcordance rate of 7.0% between phenotype and genotype for clinically significant RBC antigens within reagent RBC donors; design and validation of a high-resolution meltcurve (HRM) analysis assay and a matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) assay for RBC genotyping; identification of the genetic basis of the SARA orphan antigen utilising massively parallel sequencing; and development of a novel assay utilising calibrated microspheres to determine Lan phenotype. Overall, this thesis demonstrated the utility of SNV-based genotyping or novel and emerging approaches such as antigen quantitation by flow cytometry and MPS, in providing sensitively characterised RBCs in the modern transfusion medicine laboratory.