利用三代测序技术鉴定c.586T>C突变导致的Bel亚型

Identify the Bel Subtype of C.586T>C Mutation by Third-Generation Sequencing Technology

目的利用PacBio三代测序技术鉴定c.586T>C突变导致的Bel亚型,并分析Bel亚型家系的血型血清学特点。方法ABO血型表型检测使用血清学方法,对ABO正反定型不符的家系标本进行ABO基因第6、7号外显子Sanger测序。利用PacBio三代技术对先证者及其子女的3例样本进行ABO基因全长单体型分析。结果先证者血清学表型为Bel亚型,测序技术鉴定ABO基因序列第7外显子存在c.586T>C位点突变,三代测序单体型鉴定先证者ABO基因型为ABO^(*)BEL(c.586T>C)/O01.02,其长子和长女基因型为:ABO^(*)A1.02/BEL(c.586T>C)、ABO^(*)B.01/BEL(c.586T>C)。结论c.586T>C位点突变是导致本例Bel的分子基础,PacBio三代测序技术能够精准鉴定ABO亚型单体型。

Objective To identify the Bel subtype of c.586T>C mutation by third-generation technology,and analyze the serological characteristics of the Bel subtype blood group system.Methods The ABO blood type phenotyping examination relies on serological methods.In cases where there is a discrepancy between the ABO forward and reverse typing in a familial sample,the ABO gene's 6th and 7th exons are sequenced using Sanger sequencing.The full-length haplotype analysis of ABO gene was performed in 3 samples of proband and her children using the PacBio third-generation technology.Results The proband's serological phenotype is identified as the Bel subtype.Sequence analysis reveals a mutation at position c.586T>C in the 7th exon of the ABO gene.The haplotype analysis through third-generation technology confirms the proband's ABO genotypes as ABO^(*)BEL(c.586T>C)/O01.02.As for their eldest son and daughter,their genotypes are determined as ABO^(*)A1.02/BEL(c.586T>C)and ABO^(*)B.01/BEL(c.586T>C),respectively.Conclusion The c.586T>C mutation at the site is responsible for the molecular basis of the Bel phenotype in this case.PacBio third-generation technology enables accurate identification of ABO subtype haplotypes.