Lilić Marko, Jaklin Gordana, Gojčeta Koraljka, Raos Mirela, Golubić-Ćepulić Branka
Department of Biology, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia.
School of Medicine, University of Mostar, Mostar, Bosnia and Herzegovina.
Transfusion. 2025 Jul;65(7):1319-1327. doi: 10.1111/trf.18294. Epub 2025 May 27.
Some D variants, especially those expressed by hybrid alleles, can react falsely negative in routine serology tests. Recently, an increasing number of facilities have strived to implement a method for RHD molecular screening in blood donors.
The participants in this two-year prospective study were 735 unrelated voluntary blood donors from northwestern Croatia. Blood donors were individually tested for RHD sequences by qPCR. In reactive samples, D antigen was confirmed by adsorption/elution.
RHD screening was performed by examination of RHD exons 3, 5, and 10 in 589/704 (83.7%) serologically D-negative ccee samples, all being negative. Serology results of the Rh phenotype (C+ and/or E+) were confirmed by qPCR in the remaining 115 donors (16.3%). In this group, 112 donors were RHD-negative. RHD DEL alleles were determined in two Ccee donors (0.28%): RHDDEL32 and RHDDEL44. One donor (0.14%) was homozygous for RHD deletion but revealed an RHD sequence in 5'-UTR within the RHCE locus, c.-132A. All three donors with Rh variants expressed D epitopes as displayed by adsorption/elution. The serologically weak D group (31 participants) contained seven RHD alleles.
This study reports the DEL phenotype of RHDDEL44 expression. The hybrid allele RHCED(1)-CE was described previously, but here we illustrate the presence of D epitope(s) in a donor with homozygous RHD deletion. Weak D type 40 haplotype association was elucidated. Blood donor molecular Rh testing can prevent the immunization of genuine D-negative patients by D-positive units that reacted negatively in routine serological testing.
一些D变异体，尤其是那些由杂交等位基因表达的变异体，在常规血清学检测中可能出现假阴性反应。最近，越来越多的机构致力于在献血者中实施RHD分子筛查方法。
这项为期两年的前瞻性研究的参与者是来自克罗地亚西北部的735名无血缘关系的自愿献血者。通过qPCR对献血者个体进行RHD序列检测。在反应性样本中，通过吸附/洗脱确认D抗原。
在589/704（83.7%）血清学检测为D阴性的ccee样本中，通过检测RHD外显子3、5和10进行RHD筛查，所有样本均为阴性。其余115名献血者（16.3%）的Rh表型（C+和/或E+）血清学结果通过qPCR得到证实。在该组中，112名献血者为RHD阴性。在两名Ccee献血者（0.28%）中确定了RHD DEL等位基因：RHDDEL32和RHDDEL44。一名献血者（0.14%）为RHD缺失纯合子，但在RHCE基因座的5'-UTR中发现了一个RHD序列，即c.-132A。所有三名具有Rh变异体的献血者通过吸附/洗脱显示出D表位。血清学弱D组（31名参与者）包含7个RHD等位基因。
本研究报告了RHDDEL44表达的DEL表型。之前已描述过杂交等位基因RHCED(1)-CE，但在此我们展示了一名RHD缺失纯合子献血者中存在D表位。阐明了弱D型40单倍型关联。献血者分子Rh检测可防止真正的D阴性患者被在常规血清学检测中反应为阴性的D阳性单位免疫。
