利用PacBio三代测序技术鉴定1例B/O血型嵌合体  

B/O blood group chimera identified by PacBio third-generation sequencing:a case report

作　　者：李蕊蕊 崔丛丛 郝萧 LI Ruirui;CUI Congcong;HAO Xiao(Jinan Blood Center,Jinan 250000,China)

机构地区：[1]济南市血液供保中心,山东济南250000

出　　处：《中国输血杂志》2025年第3期421-425,共5页Chinese Journal of Blood Transfusion

摘　　要：目的 利用PacBio三代测序技术对1例ABO血型正、反定型不符的患者标本进行检测,以明确其血型,探讨血清学方法与分子生物学方法在确定嵌合体血型中的应用。方法 血型血清学试验方法;PCR基因扩增ABO基因第1~7号外显子并进行Sanger法测序;PacBio三代测序技术对标本进行ABO基因全长测序及单倍型分析;短串联重复序列(short tandem repeat, STR)分型。结果 患者血型血清学结果疑为B亚型(正定型抗-B:2+mf);Sanger测序结果显示ABO基因第6外显子c.261delG,为ABO^(*)O.01.01纯合型。PacBio三代测序结果显示该患者基因型为ABO^(*)O.01.01/ABO^(*)O.01.01及低比例的ABO^(*)B.01。STR检测结果显示患者20个STR位点中有9个位点显示出3~4种分型,证实其为嵌合体。结论 该患者为B/O血型嵌合体。低比例的ABO^(*)B.01嵌合是导致血清学抗-B混合视野的真正原因。PacBio三代测序技术不仅可以确定ABO基因单倍型还可以检出低比例ABO血型中基因嵌合。Objective To determine the blood group of a patient with ABO forward and reverse typing discrepancies u-sing PacBio third-generation sequencing(TGS)technology,and to explore the application of serological methods and molecu-lar biological methods in identifying chimeric blood groups.Methods The blood group serology testing was utilized.PCR amplification and Sanger sequencing of exons 1-7 of the ABO gene were conducted.The full-length sequencing of the ABO gene and haplotype analysis were carried out by PacBio third-generation sequencing technology.Short tandem repeat typing was also performed.Results Serological testing suggested a suspected B subtype,which appeared mixed field of vision with anti-B antibodies and showed 2+mf strength agglutination.Sanger sequencing revealed a homozygous ABO^(*)O.01.01 geno-type with the c.261delG mutation in exon 6.PacBio TGS identified a predominant ABO^(*)O.01.01/ABO^(*)O.01.01 genotype and a low proportion of ABO^(*)B.01.Nine locis of twenty short tandem repeat(STR)locis showed three or four types of gen-otypes in STR analysis,confirming chimerism.Conclusion The sample was a B/O blood group chimerism.The low pro-portion of ABO^(*)B.01 chimerism was the true cause for the serological mixed field of vision.The PacBio third-generation se-quencing technology can not only determine the ABO gene haplotype but also detect a low proportion gene chimerism in ABO blood groups.

关 键 词：ABO正反不符 嵌合体 Sanger法测序 PacBio三代测序技术 血型血清学 

分 类 号：R457.11[医药卫生—治疗学]