大连RhD阴性献血者中DEL血型的分子生物学背景研究

Molecular biologicalstudy of DEL in RhD-negative blood donors, Dalian

目的研究大连地区RhD阴性献血者中,DEL血型的频率、Rh表型及DEL血型的分子生物学背景。方法收集大连市血液中心2018年11月~2019年10月,经RhD确认实验为RhD阴性的献血者标本共355份。用单克隆抗体的血清学方法检测标本的Rh C、c、E和e表型;吸收放散实验检测DEL血型;磁珠分离法提取全血DNA;熔解曲线分析法检测DEL血型标本是否存在RHD 1227G>A基因突变;Sanger测序法对RHD基因所有外显子进行测序。结果 355例RhD确认阴性的献血者中,55例(15.5%)为DEL血型,其余300例(84.5%)为RhD真阴性。在55例DEL血型中,Rh表型以Ccee为主(45/55,81.8%);而在300例RhD真阴性中,Rh表型则以ccee为主(210/300,70.0%)。55例DEL血型中,51例(92.7%)存在RHD 1227G>A基因突变,其余4例(7.3%)为RHD基因其它位点突变。结论大连地区RhD确认阴性的献血者中,DEL血型频率为15.5%,DEL血型主要以Ccee表型及RHD 1227G>A基因突变为主。

Objective To study the frequency, Rh phenotypes and molecular & biological background of D-elute(D;) phenotype in RhD-negative blood donors in Dalian. Methods A total of 355 serologically RhD-negative samples between November, 2018 and October, 2019 in Dalian Blood Center were collected, and tested for RhC,c, E,e phenotypes using monoclonal antibodies and anti-D adsorption/elution test. DNA was extracted by magnetic bead selection. RHD 1227 G>A mutation was detected by melting curve analysis. All RHD exons were sequenced by Sanger sequencing. Results Among 355 serologically RhD-negative blood donors, 55(15.5%) were identified as D;and the remaining 300 cases(84.5%) were true RhD negative. Ccee(45/55,81.8%) was the predominant Rh phenotype among 55 D;cases while ccee(210/300,70.0%) was the most prevalent Rh phenotypes in 300 true RhD negative cases. In 55 D;cases, 51(92.7%) had RHD 1227 G>A mutation, and the other 4 cases(7.3%) had mutations in other sites. Conclusion The frequency of D;was 15.5% in serologically RhD-negative blood donors in Dalian, with Ccee being the most prevalent Rh phenotype and RHD 1227 G>A the most common gene mutation.