乌鲁木齐地区23例ABO亚型的三代测序分子遗传分析及新等位基因鉴定OA

Objective To investigate the molecular genetic background of ABO subgroups among 23 multi-ethnic volun-tary unpaid blood donors in Urumqi.Methods A total of 23 blood donor samples of ABO subgroups with ABO group dis-crepancies detected by serological tube test were collected.Preliminary ABO genotyping was performed by polymerase chain reaction with sequence-specific primers(PCR-SSP)or real-time fluorescence polymerase chain reaction(RT-PCR).PacBio third-generation sequencing(TGS)was then applied for full-length ABO gene sequencing and haplotype analysis for all samples.Results A total of 46 haplotype sequences containing single nucleotide variants(SNVs)of ABO alleles were obtained from 23 samples by TGS.Common known ABO alleles were identified including ABO*A1.02,ABO*B.01,ABO*O.01.01,ABO*O.01.02 and ABO*O.01.02.Known ABO subtype alleles included ABO*A2.01,ABO*AW.37,ABO*AW.25,ABO*AW.06,ABO*B3.01,ABO*BW.07 and ABO*BW34.Three novel ABO alleles were identified:ABO*A2.NEW(c.1040_1041insA),ABO*B.NEW(c.949A>G)and ABO*B.NEW(c.1029G>A),all of which have been deposi-ted and released in NCBI GenBank.Conclusion Third-generation sequencing can accurately obtain the complete sequence of ABO gene and distinguish haplotypes.This study elucidates the molecular genetic background of 23 ABO subtypes,provi-ding a reference for accurate blood group typing and ensuring transfusion safety.