PDF(Pubmed)
Abstract：
The Rh system, including the highly immunogenic D antigen, is one of the clinically most important blood group systems in transfusion medicine. Numerous alleles of the RHD gene are associated with variant RhD phenotypes. In case of Rh incompatibility, some of them can induce hemolytic transfusion reactions and hemolytic disease of the fetus and newborn. Thus, accurate blood group diagnostics are critical for safe transfusion therapy. We characterized phenotypes of four individuals revealing weakened D expression during routine pre-transfusion testing. Standard gel card matrix techniques with monoclonal and polyclonal anti-D antibodies were used for serological typing, complemented using D epitope and antigen density analysis. Genotyping employing PCR with sequence-specific primers, genomic and allele-specific Sanger sequencing and in silico protein analysis were performed. Four novel RHD alleles associated with weak D or partial D phenotypes were identified. One of the mutations is predicted to disrupt the terminal stop codon and result in an elongated translation of the mutant D protein that phenotypically exhibits a loss of D epitopes. Furthermore, a hybrid gene formed with the homologue RHCE gene is described. The presented data enhances the understanding of the Rh system and may contribute to continued advances in blood group diagnostics.
摘要：
Rh系统,包括高免疫原性的D抗原,是输血医学中临床上最重要的血型系统之一。RHD基因的许多等位基因与变异的RhD表型相关。在Rh不相容的情况下，其中一些会引起胎儿和新生儿的溶血性输血反应和溶血病。因此，准确的血型诊断对于安全的输血治疗至关重要.我们对四个个体的表型进行了表征，这些个体在常规输血前测试中显示出D表达减弱。使用具有单克隆和多克隆抗D抗体的标准凝胶卡基质技术进行血清学分型，使用D表位和抗原密度分析进行补充。使用序列特异性引物的PCR进行基因分型，进行了基因组和等位基因特异性Sanger测序和计算机蛋白质分析.鉴定了与弱D或部分D表型相关的四个新的RHD等位基因。预测突变之一破坏末端终止密码子并导致表型上显示D表位丢失的突变D蛋白的延长翻译。此外，描述了与同源RHCE基因形成的杂合基因。所提供的数据增强了对Rh系统的理解，并可能有助于血型诊断的不断进步。
