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目的探讨等候肾移植患者中RHD阴性血型基因血清学与分子生物学的差异。方法收集2006年1月~2016年1月广州市各器官移植中心等候肾移植的患者中经血清学检测为Rh D阴性的血样103例,采用分子生物学方法(定量PCR-测序技术)进行RHD基因分型。血清学与分子生物学两种方法对Rh D假阴性检出率行χ2检验。结果 103例血样中,Rh D真阴性(即10个外显子全部缺失)56例(54.5%),Rh D假阴性(即Rh D阳性,但是10个外显子中有部分缺失、重复、错义突变)47例(45.6%)。在47例Rh D假阴性中,弱D(weak D)1例(2.1%)、部分D(partial D)13例(27.7%)、放散D(D-elution)33例(70.2%)。血清学与分子生物学对Rh D阴性识别的差异呈统计学显著性(P<0.05)。结论 103例血清学检测出的Rh D阴性中有45.6%通过分子生物学方法检测实际为Rh D阳性变异体,说明血清学技术对识别Rh D阴性呈较高错误率。利用分子生物学技术进行受者和供者RHD血型基因分型,对于精确肾移植配型具有重要的临床意义。
Objective To investigate the serological and molecular biology differences of RHD-negative blood group genes in patients waiting for kidney transplantation. Methods 103 patients with seronegative Rh-negative blood samples from patients receiving kidney transplants at various organ transplant centers in Guangzhou from January 2006 to January 2016 were enrolled in this study. Molecular biology (quantitative PCR-sequencing) Genotyping. Two methods of serology and molecular biology Rh D false negative detection rate of χ2 test. Results Of the 103 blood samples, 56 (54.5%) had Rh D true negative (ie, all 10 exons were deleted), Rh D false negative (ie, Rh D positive but some of 10 exons were missing, Missense mutation) 47 cases (45.6%). Among 47 Rh D false negatives, weak D (2.1%), partial D 13 (27.7%) and D-elution 33 (70.2%) were weak D. Serological and molecular biology of Rh D-negative differences were statistically significant (P <0.05). Conclusions 45.6% of the 103 RhD-negative serological tests were actually RhD positive by molecular biology method, indicating that the serological technique had a higher error rate in identifying negative Rh D. The use of molecular biology techniques for RHD blood grouping of recipients and donors has important clinical implications for accurate kidney transplantation.