RHD and RHCE molecular analysis in weak D blood donors and in patients with Rh antibodies against their own corresponding Rh antigen.

backgroundThe Rh system is the largest and most polymorphic blood group system. The existence of a large number of RH alleles results in variant phenotypes that often complicate blood donor phenotyping and the distinction between auto- and allo-antibodies in recipients who have anti-Rh antibodies in the presence of their own corresponding Rh antigen. Knowledge of these variants is necessary in order to make blood transfusion safer. MATERIALS AND

methodsSamples from 48 blood donors with serological weak D and from 29 patients who had anti-Rh antibody in the presence of their own corresponding Rh antigen were evaluated molecularly for RHD and RHCE alleles using a blood-multiplex ligation-dependent probe amplification assay and Sanger sequencing.

resultsRh variants were found in 45 of the 48 blood donors: 24/45 (53%) were weak D, 2/45 (4%) partial D and 19/45 (42%) were weak and partial D. The remaining three donors (6%) did not show a mutation in the RHD allele. Among the 29 patients, 13/29 had anti-e, of whom 4/13 had genotypes that predicted a partial e antigen; 11/29 had anti-D, with 6/11 being identified as partial D; 2/29 had anti-c, of whom 1/2 was predicted to express partial c antigen; 4/29 who had anti-E and 4/29 who had anti-C did not show mutations in RHCE*C or RHCE*E. DISCUSSION: It was possible to find individuals with clinically significant Rh phenotypes due to the weak reactivity of the D antigen, detected through serological tests in blood donors. In patients, when found with the anti-Rh antibody in the presence of the same Rh antigen, it is difficult to distinguish an auto-antibody from an allo-antibody by serological tests; in these cases, molecular methods (genotyping) can help us to determine whether there are changes in the RH alleles and to discover the nature of the antibody (allo or auto).