Legler T J, Maas J H, Köhler M, Wagner T, Daniels G L, Perco P, Panzer S
Department of Transfusion Medicine, University of Göttingen, Robert-Koch-Str. 40, 37075 Göttingen, Germany.
Transfus Med. 2001 Oct;11(5):383-8. doi: 10.1046/j.1365-3148.2001.00327.x.
The serological differentiation of weak D from partial D, D-negative and D-positive is not always unequivocal. Therefore, sequencing of the RHD gene is required in some cases. Very recently, several new differences between RHD and RHCE have been identified which permitted us to design primers close to the exon/intron boundaries of the RHD-exons. We evaluated these primers in 83 D-positive and 18 D-negative blood donors and applied the new method for the characterization of the RHD gene in six individuals with weak D phenotype. The amplification reactions were concordant with serological findings in 100 of 101 donors (99.0%). In one D-positive donor the PCR for exons 2 and 5 gave a negative result, while the sequence of the remaining eight exons was unchanged. By sequencing samples with very weak D serological reactions, we identified weak D type 4.2.2 and weak D type 15, both previously reported to be associated with anti-D-alloimmunization. Consequently, we recommended the selection of D-negative blood in the weak D type 4.2.2 patient, and the provision of Rh prophylaxis for pregnant women with weak D type 15. In summary, a new RHD sequencing method was developed which can be applied if serological reactions are inconclusive.
弱D与部分D、D阴性及D阳性的血清学鉴别并不总是明确的。因此，在某些情况下需要对RHD基因进行测序。最近，已经确定了RHD和RHCE之间的几个新差异，这使我们能够设计靠近RHD外显子外显子/内含子边界的引物。我们在83名D阳性和18名D阴性献血者中评估了这些引物，并将这种新方法应用于6名弱D表型个体的RHD基因特征分析。101名献血者中有100名（99.0%）的扩增反应与血清学结果一致。在一名D阳性献血者中，外显子2和5的PCR结果为阴性，而其余八个外显子的序列未改变。通过对血清学反应非常弱的样本进行测序，我们鉴定出了弱D 4.2.2型和弱D 15型，这两种类型此前均被报道与抗-D同种免疫有关。因此，我们建议为弱D 4.2.2型患者选择D阴性血液，并为弱D 15型孕妇提供Rh预防措施。总之，开发了一种新的RHD测序方法，当血清学反应不确定时可应用该方法。
