Faas B H, Simsek S, Bleeker P M, Overbeeke M A, Cuijpers H T, von dem Borne A E, van der Schoot C E
Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam.
Blood. 1995 Feb 1;85(3):829-32.
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.
已表明恒河猴（Rh）血型抗原由两个同源基因编码：Rh D基因和Rh CcEe基因。Rh CcEe基因编码不同的肽：Rh C、c、E和e多肽。在编码Rh E和Rh e抗原多肽的等位基因之间仅发现一个核苷酸差异。它是核苷酸位置676处的C→G转换，这导致携带Rh e的多肽中脯氨酸被丙氨酸取代。在此，我们提出一种等位基因特异性引物扩增（ASPA）方法来确定Rh E和Rh e基因型。在一次聚合酶链反应中，正向引物的3'端核苷酸对Rh E等位基因位置676处的胞嘧啶具有特异性。在另一次反应中，使用的正向引物其3'端核苷酸对Rh e等位基因和Rh D基因位置676处的鸟嘌呤具有特异性，而反向引物的3'端核苷酸对Rh CcEe基因位置787处的腺嘌呤具有特异性。我们在这些检测中测试了来自158名正常供体（包括非白种人供体和具有罕见Rh表型的供体）的DNA样本。结果与血清学Rh E/e表型分析结果完全一致。因此，我们可以得出结论，ASPA方法导致一种简单可靠的方法来确定Rh E/e基因型。这在胎儿的Rh E/e基因分型和/或在没有红细胞可用于血清分型的情况下可能有用。此外，我们的结果证实了位置676处的胞嘧啶/鸟嘌呤多态性与Rh E/e表型之间的关联。
