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.
题目: RhC抗原弱表达伴类自身抗-Ce及同种抗-Jkb致配血不合分析.
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。.
Keywords: genetic recombination; antibody screening; mimicking anti-Ce; anti-Jkb.