1例RH血型基因缺失患者及家系的实验性研究  

An experimental study on a patient with RH blood group gene deletion and his pedigree

作　　者：吴争胜[1] 杜垚强 陈秉宇[1] 王彬[1] WU Zheng-sheng;DU Yao-qiang;CHEN Bing-yu;WANG Bin(Department of Transfusion Medicine,Zhejiang Provincial People's Hospital,Affiliated People's Hospital of Hangzhou Medical College,Hangzhou,Zhejiang 310014,China)

机构地区：[1]浙江省人民医院杭州医学院附属人民医院输血科,浙江杭州310014

出　　处：《中国卫生检验杂志》2022年第2期135-138,共4页Chinese Journal of Health Laboratory Technology

基　　金：教育部科技发展中心基金项目(2017A11036);浙江省自然科学基金项目(LQ21H200007)。

摘　　要：目的研究RH基因缺失型表型患者及家系的血型血清学特征,分析其表型基因序列的家系遗传背景和临床意义。方法采集患者及其家属的EDTA-K2抗凝血,进行血型血清学检测标本RHD、C、c、E、e抗原,同时提取标本DNA,采用RHD和RHCE血型分型试剂盒及荧光定量PCR分型,然后进行RHD和RHCE血型测序试剂盒加样、PCR扩增,产物电泳切胶,上机Sanger测序。结果患者血清学检测B型RH D阳性,缺失C、c、E、e抗原,直接抗人球蛋白阴性,抗体鉴定16谱细胞阳性,交叉配血与献血员均不相合。RHD、RHCE基因分子生物学鉴定得到,患者RHCE基因的缺失由RHD替换,患者基因型CE-D(2-9)-CE//Null,表型D--,产生抗体极可能为抗-Hr0;其配偶基因型CE//Ce,表型CCDEe;大女儿基因型CE//Null,表型CDE;小女儿基因型CE//CE-D(2-9)-CE,表型CDE。结论RHCE基因容易与RHD基因存在重叠交叉,以及基因突变,导致RH表型缺失,中国人群中存在一定比例的抗-Hr0,临床输血需要引起高度重视。Objective To study the blood group serological characteristics of a patient with RH gene deletion phenotype and his pedigree,and to analyze the family genetic background and clinical significance of the phenotypic gene sequence.Methods EDTA anticoagulant blood samples from patients and their families were collected for RHD,C,c,E and e antigens of blood group serological test samples.Meanwhile,DNA of the samples were extracted and typed by RHD and RHCE blood group typing kits and fluorescence quantitative PCR.Samples were added and PCR amplified by RHD and RHCE blood group sequencing kits.Then we performed the Sanger sequencing in each samples.Results The serological test of the patient was positive for type B RHD+,missing C,c,E,and e antigens,negative for direct anti-screening,positive for anti-screen 16 spectrum,and inconsistent cross-matching.Molecular biological identification of RHD and RHCE genes was performed.The deletion of RHCE in the patient was replaced by RHD.The patient's genotype was CE-D(2-9)-CE//Null,phenotype D--,and the antibody produced was most likely anti-Hr0;His spouse's genotype was CE//Ce,phenotype was CCDEe;Elder daughter's genotype was CE//Null,phenotype was CDE;Younger daughter's genotype was CE//CE-D(2-9)-CE,phenotype was CDE.Conclusion The RHCE gene is prone to overlap with the RHD gene,as well as gene mutations,resulting in the loss of the RH phenotype.There is a certain proportion of anti-Hr0 in the Chinese population,and clinical blood transfusion should pay more attention to this situation.

关 键 词：RHD RHCE 基因缺失 荧光定量PCR 测序分析 

分 类 号：R392.11[医药卫生—免疫学]