1例含RHD基因的RhD阴性表现型分析

A Case Report of RhD Positive Genotye with RhD Negative Phenotype

目的分析1例RhD阴性表现型而RHD基因分型阳性的献血者样本,了解其发生的分子基础。方法采用微量板法对献血者进行RhD阴性筛查,采用抗人球蛋白法对初筛RhD阴性样本进行确认,从静脉血中提取献血者基因组DNA,采用PCR-SSP法进行RHD基因分型,并对RHD基因进行直接测序。结果经抗人球蛋白法确认33例RhD阴性献血者中,发现RHD阳性基因型1例。对该例样本测序分析,发现其RHD基因外显子5存在纯合的基因变异,即在744的位置插入了AG,同时删除了GTGGTGACAGCCATC 15个碱基。结论利用PCR-SSP法对血清学RhD阴性样本进行基因型检测,并结合RHD基因测序分析,更能准确鉴定RhD血型。

Objective To analyze a blood donor of RhD negative phenotype with RhD positive genotype.Methods Microplating method was used to screen RhD negative samples among blood donors and anti-globulin method was used to confirm RhD negative phenotype.Genomic DNA was extracted from peripheral blood.PCR-SSP method was used to identify the RhD genotype.Additionally,the coding regions of RhD were analyzed by DNA sequencing.Results One case with RhD positive genotype was found among 33 RhD negative samples that were confirmed by anti-globulin method.RhD gene sequencing revealed homozygous combination mutation in exon 5 （AG insertion combined with 15 nucleotides of GTGGTG ACAGCCATC deletion located at 744 bp）.Conclusion PCR-SSP method combined with RhD gene sequencing may improve the accuracy of RhD blood group identification for the serologically identified RhD negative samples.