郑州地区RhD变异型献血者分子生物学分析

Molecular biological analysis of RhD variant blood donors in Zhengzhou

目的研究郑州地区RhD变异型献血者血清学特征及基因突变机制。方法选择2023年1月—2023年12月送至本实验室的1619名RhD阴性献血者为研究对象,应用试管法和微柱凝胶间接抗球蛋白试验方法进行RhD阴性确认试验及RhCE表型检测。采用RHD基因扩增和Sanger测序检测RhD变异型标本基因型。结果RhD阴性确认试验共检出RhD变异型69例,比例为4.26%(69/1619)。RhCE表型分别为ccEe、Ccee、CcEe、CCee。包含17种基因型,15种D变异型表型。RHD^(*)weak partial 15等位基因最多(33例),频率为47.83%(33/69),主要表型为ccEe;其次是RHD^(*)DVI.3等位基因20例,频率为28.99%(20/69),主要表型为Ccee。RHD^(*)weak partial 15/RHD^(*)01EL.01杂合子3例,频率为4.35%(3/69),皆为CcEe表型。其他罕见基因型共13例,频率为18.84%(13/69)。抗筛阳性3例,比例为4.35%(3/69)。女性献血者2名,皆有孕产史,经抗体鉴定为抗-D;男性献血者1名,为抗-M。结论郑州地区RhD变异型献血者中RHD^(*)weak partial 15基因型最常见,其次为RHD^(*)DVI.3基因型。对保障安全供血,指导临床精准输血具有重要作用。

Objective To investigate the serological characteristics and gene mutation mechanism of RhD variant blood donors in Zhengzhou.Methods From January 2023 to December 2023,1619 RhD-negative blood donors sent to our laboratory were selected for the study,and RhD negative confirmation test and RhCE phenotype detection were applied by tube method and microcolumn gel indirect antiglobulin test method.RHD gene amplification and Sanger sequencing were used to detect RhD variant sample genotypes.Results A total of 69 cases of RhD variants were detected in the RhD negative confirmation test,with a proportion of 4.26%(69/1619).The RhCE phenotypes were ccEe,Ccee,CcEe and CCee.There were 17 genotypes and 15 phenotypes of the D variant.The RHD^(*)weak partial 15 allele was the most frequent(33 cases),with a frequency of 47.83%(33/69),and the main phenotype was the ccEe.This was followed by the RHD^(*)DVI.3 allele in 20 cases with a frequency of 28.99%(20/69)and the predominant phenotype was Ccee.The RHD^(*)weak partial 15/RHD^(*)01EL.01 heterozygote was found in 3 cases with a frequency of 4.35%(3/69),all with the CcEe phenotype.Other rare genotypes were present in 13 cases with a frequency of 18.84%(13/69).Antibody screening was positive in 3 cases with a frequency of 4.35%(3/69).Two cases of female blood donors,both with history of pregnancy and childbirth,were identified as anti-D;one case of male donor was anti-M.Conclusion The RHD^(*)weak partial 15 genotype was the most common among the RhD variants in blood donors in Zhengzhou,followed by the RHD^(*)DVI.3 genotype.It plays an important role in guaranteeing the safety of blood supply and guiding precision transfusion.