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BH Faas, S Simsek, PM Bleeker, MA Overbeeke, HT Cuijpers, AE von dem Borne and CE van der Schoot
Central Laboratory of the Netherlands Red Cross Blood Transfusion Service,
Amsterdam.
It has been shown that the Rhesus (Rh) blood group antigens are encoded by
two homologous genes: the Rh D gene and the Rh CcEe gene. The Rh CcEe gene
encodes different peptides: the Rh C, c, E, and e polypeptides. Only one
nucleotide difference has been found between the alleles encoding the Rh E
and the Rh e antigen polypeptides. It is a C-- >G transition at
nucleotide position 676, which leads to an amino acid substitution from
proline to alanine in the Rh e-carrying polypeptide. Here we present an
allele-specific primer amplification (ASPA) method to determine the Rh E
and Rh e genotypes. In one polymerase chain reaction, 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 primer was used with a 3'-end
nucleotide specific for the guanine at position 676 of the Rh e allele and
the Rh D gene, whereas the antisense primer 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 (including non-Caucasian donors and donors
with rare Rh phenotypes) in these assays. There was full concordance with
the results of serologic Rh E/e phenotyping. Thus, we may conclude that the
ASPA approach leads to 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 results confirm the proposed association between the cytosine/guanine
polymorphism at position 676 and the Rh E/e phenotype.
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