Hou Jue, Xia Yu, Li Jian, Chen Xue, Li Meng, Zhao Yuwei, Fu Xuemei
Blood Grouping Reference Laboratory, Chengdu Blood Center, Sichuan, China.
Blood Grouping Reference Laboratory, Chengdu Blood Center, Sichuan, China.
Transfus Clin Biol. 2025 Jun 16. doi: 10.1016/j.tracli.2025.06.002.
Some D variant individuals are at risk of developing anti-D immunization when exposed to RhD-positive red cells. Therefore, accurate typing of D variants is essential to ensure the safety of clinical blood transfusion practices. The aim of this study was to investigate the serological profiles and molecular characteristics of D variants among Chengdu blood donors and to establish a more cost-effective RHD genotyping strategy tailored for the Chinese population.
During the period from 2019 to 2022, samples from Chengdu blood donors typed as RhD-negative with IgM anti-D using the microplate method underwent additional RhD typing with a panel of monoclonal anti-D reagents using both the micro-column gel card technique and the saline tube method. Samples that could not be classified as either RhD-positive or RhD-negative were genotyped using the PCR-SSP assay and Sanger sequencing. RHD zygosity status was determined by assessing the presence or absence of hybrid Rhesus boxes. Alloantibody screening was conducted to evaluate the risk of anti-D immunization.
Three variants RHD15, RHDDEL1, and RHD06.03.01 accounted for 61% of D variants identified in our study. Each of these were studied for distinct reaction patterns with a panel of anti-Ds. Additionally, eight previously reported D variant alleles, including RHD01W.95, RHD01W.72, RHD01W.12, RHD01W.18, RHD01W.39, RHD01W.67, RHD01W.71, and RHD*01W.960A were observed sporadically. Furthermore, two novel alleles characterized by a nucleotide change (c.283G>T) and a nucleotide change (c.84C>G) respectively were identified. No cases of anti-D formation were detected in the D variant samples.
The spectrum of D variants identified in this study highlights the genetic diversity in the Chengdu blood donors, underscoring the need to incorporate these D variants into RHD genotyping strategy tailored specifically for the Chinese population. Comprehensive analysis of serological reaction patterns across these D variants would help guide molecular testing strategies and enhance genotyping cost-efficiency.
部分D变异个体在接触RhD阳性红细胞时存在产生抗-D免疫的风险。因此，准确鉴定D变异体对于确保临床输血操作的安全性至关重要。本研究旨在调查成都献血者中D变异体的血清学特征和分子特征，并建立一种更具成本效益的、针对中国人群的RHD基因分型策略。
在2019年至2022年期间，对采用微孔板法用IgM抗-D分型为RhD阴性的成都献血者样本，使用一组单克隆抗-D试剂，采用微柱凝胶卡技术和盐水试管法进行额外的RhD分型。无法归类为RhD阳性或RhD阴性的样本采用PCR-SSP检测和桑格测序进行基因分型。通过评估杂交恒河猴盒的存在与否来确定RHD纯合性状态。进行同种抗体筛查以评估抗-D免疫的风险。
三种变异体RHD15、RHDDEL1和RHD06.03.01占本研究中鉴定出的D变异体的61%。对其中每一种变异体与一组抗-D的不同反应模式进行了研究。此外，还偶尔观察到八个先前报道的D变异等位基因，包括RHD01W.95、RHD01W.72、RHD01W.12、RHD01W.18、RHD01W.39、RHD01W.67、RHD01W.71和RHD*01W.960A。此外，还鉴定出两个分别以核苷酸变化（c.283G>T）和核苷酸变化（c.84C>G）为特征的新等位基因。在D变异体样本中未检测到抗-D形成的病例。
本研究中鉴定出的D变异体谱突出了成都献血者中的基因多样性，强调需要将这些D变异体纳入专门针对中国人群的RHD基因分型策略。对这些D变异体的血清学反应模式进行综合分析将有助于指导分子检测策略并提高基因分型的成本效益。
