RH E E GENOTYPING BY ALLELE-SPECIFIC PRIMER AMPLIFICATION/

It has been shown that the Rhesus (Rh) blood group antigens are encode d by two homologous genes: the ph D gene and the Rh CcEe gene. The Rh CcEe gene encodes different peptides: the Rh C, c, E, and e polypeptid es. Only one nucleotide difference has been found between the alleles encoding the Rh E and the ph e antigen polypeptides. It is a C --> G t ransition at nucleotide position 676, which leads to an amino acid sub stitution from proline to alanine in the Rh e-carrying polypeptide. He re we present an allele-specific primer amplification (ASPA) method to determine the Rh E and Rh e genotypes. In one polymerase chain reacti on, the sense primer had a 3'-end nucleotide specific for the cytosine at position 676 of the Rh E allele. In another reaction, a sense prim er was used with a 3'-end nucleotide specific for the guanine at posit ion 676 of the Rh e allele and the Rh D gene, whereas the antisense pr imer had a 3'-end nucleotide specific for the adenine at position 787 of the Rh CcEe gene. We tested DNA samples from 158 normal donors (inc luding non-Caucasian donors and donors with rare Rh phenotypes) in the se assays. There was full concordance with the results of serologic Rh E/e phenotyping. Thus, we may conclude that the ASPA approach leads t o a simple and reliable method to determine the Rh E/e genotype. This can be useful in Rh E/e genotyping of fetuses and/or in cases in which no red blood cells are available for serotyping. Moreover, our result s confirm the proposed association between the cytosine/guanine polymo rphism at position 676 and the Rh E/e phenotype. (C) 1995 by The Ameri can Society of Hematology.