135例ABO变异型的血清学特征和基因序列分析  

Serological characteristics and gene sequence analysis of ABO variants in 135 cases

作　　者：孔存权 孔永奎[2] 崔翠云 孔晓阳 朱伟彦 刘鸿雁 蒋姝婷 马翠霞 戴豫宛 燕备战 KONG Cunquan;KONG Yongkui;CUI Cuiyun;KONG Xiaoyang;ZHU Weiyan;LIU Hongyan;JIANG Shuting;MA Cuixia;DAI Yuwan;YAN Beizhan(Department of Blood Transfusion,Henan Provincial People's Hospital,Zhengzhou University People's Hospital,Zhengzhou,Henan 450003,China;Department of Blood Transfusion,the First Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450052,China)

机构地区：[1]河南省人民医院输血科郑州大学人民医院,河南郑州450003 [2]郑州大学第一附属医院输血科,河南郑州450052

出　　处：《中华实用诊断与治疗杂志》2025年第1期38-44,共7页Journal of Chinese Practical Diagnosis and Therapy

基　　金：河南省科技攻关计划项目(182102310223)。

摘　　要：目的 分析135例ABO变异型者的血清学特征和ABO血型基因型,探讨其分子生物学基础。方法 2021年1月—2024年7月河南省人民医院行ABO血型鉴定者265 025例,筛选出135例疑似ABO亚型需基因学辅助诊断。提取全血DNA后,采用PCR技术对ABO血型基因1~7外显子及其侧翼序列、调控区序列扩增和测序。应用Modeller v10.4软件针对AB_(w)亚型B链上的1043突变(p.His348Pro)构建α-1,3-半乳糖基转移酶三维分子同源模型,并通过Discovery studio 2016软件进行能量优化,使用Pymol软件进行比对分析。结果 265 025例共检出ABO亚型68例,亚型检出率约2.6/万。68例检出等位基因36个,包括5个新等位基因。A_(el)型与B_(el)型抗原表达量极弱,部分吸收放散实验仍不能检出,A_(el)型多有不规则抗A抗体,B_(el)型中多有不规则抗B抗体,血清学检测极易判定为O型。AB_(w)亚型B链同源建模显示,B.01基础上出现c.1043A>C突变导致AB_(w)型GTB的C-末端的α螺旋缩短,Pro348与邻近氨基酸之间形成氢键减少,仅剩与Arg352间1条氢键。结论 ABO亚型中B(A)型在人群中出现比例较高,B_(w)型和A_(el)型次之;血清学上A_(el)亚型与B_(el)亚型极易误定为O型;B.01基础上出现c.1043A>C突变可能影响GTB酶活性,形成B_(w)表型。Objective To analyze the serological characteristics and ABO genotypes of 135patients with ABO variants,and to explore its molecular biological basis.Methods Totally 265 025patients underwent ABO blood typing in Henan Provincial People's Hospital from January 2021to July 2024,among whom 135patients were screened as suspected ABO subtypes and were subjected to genetic auxiliary diagnosis.After the whole blood DNA was extracted,PCR technology was used to amplify and sequence ABO gene exons 1-7and their flanking and regulatory region sequences.Modeller v10.4software was used to construct a three-dimensional molecular homology model ofα-1,3-glycosyltransferase B(GTB)with1043mutation (p.His348Pro)on the B chain of AB_w.Discovery studio 2016software was performed for energy optimization.Pymol software was used for comparative analysis.Results In 265 025patients,68were detected ABO subtype,with a detection rate of approximately 2.6per 10 000.Among them,36alleles were detected including 5novel alleles.The antigen expression levels of A_(el)and B_(el )were extremely weak,and some specimens were not detected by absorption and release experiments.A_(el )mostly had irregular anti-A and B_(el )mostly had irregular anti-B,and serological tests were easily identified as O type.Homologous modeling results of the AB_(w )B chain showed that the c.1043A>C mutation on the basis of B.01 resulted in a shortened C-terminalα-helix of AB_(wtype) GTB,with reduced hydrogen bonds between Pro348and adjacent amino acids,leaving only one hydrogen bond with Arg352.Conclusions The B(A)type of ABO subtypes has a higher prevalence proportion,followed by B_(w )type and A_(el)type.Serologically,the A_(el)and B_(el )subtypes are easily misidentified as O type.The c.1043A>C mutation on the basis of B.01 may impact GTB enzyme activity and lead to B_(wphenotype) formation.

关 键 词：ABO变异型 血清学 基因型 新等位基因 同源建模 

分 类 号：R440[医药卫生—诊断学]