1例RHD-CE(3-7)-D基因重组与RHCE变异型患者的血清学与分子生物学分析  

Serological and molecular biological analysis of a patient with RHD⁃CE(3⁃7)⁃D gene recombination and RHCE*cE variant

作　　者：唐炳娣 蔡仲仁 邓泳诗 伍昌林 TANG Bingdi;CAI Zhongren;DENG Yongshi;WU Changlin(Guangdong Medical University,Zhanjiang,Guangdong,China,524023;Department of Blood Transfusion,Huizhou Third People's Hospital,Huizhou,Guangdong,China,516002;Department of Blood Transfusion,Third People's Hospital of Shenzhen Longgang District,Shenzhen,Guangdong,China,518116;Department of Blood Transfusion,Shenzhen Second People's Hospital,Shenzhen,Guangdong,China,518035)

机构地区：[1]广东医科大学,广东湛江524023 [2]惠州市第三人民医院输血科,广东惠州516002 [3]深圳市龙岗区第三人民医院输血科,广东深圳518116 [4]深圳市第二人民医院输血科,广东深圳518035

出　　处：《分子诊断与治疗杂志》2024年第6期1183-1186,1190,共5页Journal of Molecular Diagnostics and Therapy

基　　金：广东省基础与应用基础研究基金项目(2020A1515011520)。

摘　　要：目的 研究分析1例Rh血型弱D、弱cE患者的血清学与分子生物学特征,为该类患者的临床安全输血提供实验依据。方法 采用微柱凝胶卡法对患者红细胞进行ABO、RhDCcEe抗原的鉴定,同时采用试管法进行血型复核,抗人球蛋白卡法筛查不规则抗体;采用PCR-SSP法对RhDCcEe(RhD、RhC、Rhc、RhE、Rhe)基因型进行检测;三代全长测序技术对RHD/RHCE基因序列进行测序分析。结果 微柱凝胶卡法鉴定ABO、RhD、RhCcEe血型抗原的结果为:A抗原(-)、B抗原(-)、RhD(1+)、RhC(4+)、Rhc(1+)、RhE(1+)、Rhe(4+)、对照孔(-);试管法ABO、RhD、RhCcEe抗原鉴定该患者表型为:A抗原(-)、B抗原(-)、RhD(w+)、RhC(4+)、Rhc(w+)、RhE(w+)、Rhe(4+),对照管(-);抗人球蛋白卡法筛查患者不规则抗体阴性;PCR-SSP法血型基因分型RhDCcEe结果:RhD(+)、RhC(+)、Rhc(+)、RhE(+)、Rhe(+);RHD/RHCE基因结果:RHD单倍体1为外显子1-10全缺失,而单倍体2为外显子RHD-CE基因重组融合,且确认其重组类型为RHD-CE(3-7)-D,起点在外显子2(g.20238-20312之间),终点在外显子8(g49184-50480之间),同时RHCE基因第6外显子存在新碱基点突变RHCE*cE(827C>A)。结论RHD-CE(3-7)-D基因重组融合与RHCE*cE(827C>A)新等位基因突变可能引起D、cE血型抗原弱表达,为临床安全输血提供了重要的实验数据支持。Objective To investgate a case of RHD-CE(3-7)-D gene recombination and RHCE*cE variant for serological and molecular biological characteristics,to provide experimental basis for the clinical safe blood transfusion.Methods ABO,RhD,and RhCcEe antigens were identified using the micro-column gel card method.Blood group was determined using the blood test tube method,and irregular antibodies were screened using the anti-human globulin card method.The genotypes of RhD,RhC,Rhc,RhE,and Rhe were detected using PCR-SSP,and the entire sequence of the RHD/RHCE gene was analyzed using three-genera-tion full-length sequencing.Results ABO,RhD,and RhCcEe blood group antigens were tested as follows:A(-),B(-),RhD(1+),RhC(4+),Rhc(1+),RhE(1+),Rhe(4+)using a micro-column gel card.Upon repeating the test using the test tube method,the results were A(-),B(-),RhD(w+),RhC(4+),RhC(w+),RhE(w+),RhE(4+).Anti-human globulin card screening for irregular antibody was negative.Geno-typing results of RhDCcEe by PCR-SSP:RHD(+),RHC(+),RHc(+),RHE(+),RHe(+).Analysis of the RHD/RHCE gene showed that RHD haploid 1 had a complete deletion in exons 1-10,while haploid 2 exons showed a RHD-CE gene recombination,it was confirmed that its recombination type was RHD-CE(3-7)-D.The starting point of this recombination was identified in exon 2(g.20238-20312),and the end point was deter-mined in exon 8(g.49184-50480).Additionally,a new base point mutation was found in exon 6 of the RHCE gene(827C>A).Conclusion RHD-CE(3-7)-D gene recombination and RHCE*cE(827C>A)novel allele mutation may cause weak expression of D and CE blood group antigens.This provides a foundation for safe blood transfusions in clinical settings.

关 键 词：RhD/cE弱抗原 RHD-CE(3-7)-D重组 RHCE*cE(827C>A) 三代全长测序 

分 类 号：R457.1[医药卫生—治疗学]