Clinical aspects of Rh genotyping

The Rh antigens are encoded by the RHD and RHCE genes. In RhD negative individuals the RHD gene is absent or grossly deleted. Routinely, Rh typing is performed by haemagglutination. However, there are some clinical situations in which serological techniques are not suitable for determining the red blood cell phenotype accurately. Most anti-D sera may not agglutinate erythrocytes possessing a reduced expression of the D antigen. In these cases, DNA-based analyses may be better than serological typing to infer the appropriate phenotype. Agglutination methods are also of limited use for determining the red blood cell phenotype of a foetus at risk of haemolytic disease of the newborn. Molecular RHD typing using amniocytes or DNA obtained from maternal plasma may obviate the need of amniocenteses during pregnancy when the foetus is RhD negative, thus providing an important tool in managing possible sensitization by foetal erythrocytes. Classical haemagglutination has limitation in patients with autoimmune haemolytic anaemia. Erythrocytes coated with IgG cannot be accurately typed for red blood cell antigens, particularly when directly agglutinating antibodies are not available or IgG removal by chemical treatment is insufficient. Molecular genotyping is very important for determination of the true blood group antigens of these patients. RHD genotyping with a specificity and sensitivity comparable to serologic methods is of practical importance to overcome the limitations of serology and, in addition, to improve the currently possible resolution.