一例ABO血型Aw33亚型新等位基因的分子生物学鉴定

目的研究1例ABO血型A亚型的分子生物学机制。方法患者ABO血型正反定型采用卡式法和试管法分别进行鉴定。利用PCR序列特异性引物检测该患者所含ABO基因。PCR扩增该患者ABO基因1~7外显子的全部编码序列并进行测序分析,通过克隆测序进行ABO基因单倍型分析。结果患者红细胞与抗A呈现弱凝集,与抗B不凝集,其血清与Ac凝集1+,与Bc呈现4+凝集,血清学特性可定义为Aw亚型。ABO基因测序分析显示患者存在c.106G>T、c.188G>A、c.189C>T、c.220C>T、c.297A>G、c.467C>T、c.543G>C、c.646T>A、c.681G>A、c.771C>T、c.829G>A杂合变异和c.261delG缺失。结合克隆测序结果,推测患者ABO基因型为ABO*Aw.33.new/O.01.02;与ABO*A1.01相比存在c.467C>T和c.543G>C变异,与ABO*A1.02相比存在c.543G>C变异,该新等位基因序列已提交GenBank,序列号为MK302122。结论发现1例Aw33亚型新的等位基因,其GTA转移酶基因存在c.467C>T和c.543G>C变异。

Characterization of a novel allele of Aw33 subtype of the ABO blood group

Objective To explore the molecular basis for an A subtype of the ABO blood group.Methods The forward and reverse typing of the ABO blood group were identified by gel card and test tube methods.The ABO gene of the patient was detected by PCR-sequence specific primer(PCR-SSP).Exons 1 to 7 of the ABO gene was amplified by PCR and sequenced.The ABO gene was also subjected to subclone sequencing for haplotype analysis.Results The patient’s red cells showed weak agglutination with anti-A but non-agglutination with anti-B.The patient’s serum showed 1+agglutination with A cells and 4+agglutination with B cells.Based on above serological characteristics,the patient was defined as Aw subtype of the ABO blood group.Sequencing analysis showed that the patient was heterozygous for c.106G>T,c.188G>A,c.189C>T,c.220C>T,c.297A>G,c.467C>T,c.543G>C,c.646T>A,c.681G>A,c.771C>T,c.829G>A,in addition with a c.261G deletion.Combined with the result of subclone sequencing,the ABO genotype of the patient was determined as ABO*Aw.33.new/O.01.02,which harbored c.467C>T and c.543G>C variants compared with ABO*A1.01 and c.543G>C variant compared with ABO*A1.02.The novel allele has been submitted to GenBank with an accession number of MK302122.Conclusion A novel allele of Aw33 subtype has been identified with its GTA transferase gene harboring c.467C>T and c.543G>C variants compared with A1.01.