目的:探讨1例患者类自身抗-Ce合并抗-Jkb引起交叉配血不合并分析其RHCE基因弱表达的原因.方法:采用试管法、毛细管离心法对ABO、Rh和Kidd血型抗原进行鉴定.采用盐水、聚凝胺、抗人球蛋白三介质联用多套谱细胞进行抗体筛查及抗体特异性鉴定;采用多重PCR技术对RHCE基因进行测序及单倍体分析并使用Swiss-Model 进行RHCE蛋白建模.结果:该患者血清中检出类自身抗-Ce合并抗-Jkb抗体.RHCE三代单分子测序显示突变组合为 c.48G＞C、c.150C＞T、c.178C＞A、c.201A＞G、c.203A＞G 和 c.307C＞T,在内含子 2 中存在 109 bp 插入序列,同时出现内含子5-8大片段丢失,其Rh血型基因型为DCe/DCe,表型为CCDee.结论:基因分型技术可以协助推断患者血清中部分弱表达的RhC、c、E、e的分子机理,以辅助疑难抗体的鉴定,从而保证患者输血安全.
Analysis of RhC Antigen Weak Expression Combined with Mimicking Autoanti-Ce and Homologous Anti-Jkb Causing Mismatch
Objective:To investigate the reasons for the weak expression of RHCE gene in a patient whose mimicking anti-Ce combined with anti-Jkb caused cross-matching non-combination.Methods:ABO,Rh,and Kidd blood group antigens were identified by test tube method and capillary centrifugation.Antibody screening and antibody specificity identification were performed using saline,polybrene and antiglobulin in tri-media association with multispectral cells.RHCE gene sequencing and haploid analysis were performed by multiplex PCR technique and RHCE protein modeling was performed using Swiss-Model.Results:The serum of the patient contained anti-Ce mimicking autoantibodies along with anti-Jkb antibodies.c.48G＞C,c.150C＞T,c.178C＞A,c.201A＞G,c.203A＞G,and c.307C＞T mutations were detected in the RHCE triple-molecule sequencing.A 109 bp insertion sequence was found in intron 2,with fragment loss from intron 5-8.The Rh-group genotype was DCe/DCe,and phenotype was CCDee.Conclusion:Genotyping techniques can assist in deducing the molecular mechanisms of some weakly expressed RhC,c,E,and e in patients'sera to aid in the identification of difficult antibodies and thus ensure the safety of patients'blood transfusion.
万方数据
维普
