25例B亚型/AB亚型的ABO基因序列分析

Genotypic Identification and Sequence Analysis of B/AB Subtypes in 25 Cases

目的探讨25例B亚型/AB亚型标本的分子生物学机制。方法应用PCR-SSP对血型血清学确认为B抗原减弱的标本进行ABO*B基因检测或应用Sanger一代测序技术对ABO基因的第6-7外显子或第1-7外显子测序;对最终无法定型的标本进一步用PacBio:SMRT三代测序技术进行检测。结果25例B亚型/AB亚型标本均经过基因序列分析,共发现7种已知的ABO*B等位基因,分别为ABO*BW.27(3/25,12.00%)、ABO*BW.03(5/25,20.00%)、ABO*B3.03(2/25,8.00%)、ABO*BEL.03(1/25,4.00%)、ABO*BW.07(7/25,28.00%)、ABO*B3.05(1/25,4.00%)、ABO*BW.12(1/25,4.00%);5种未知的ABO*B等位基因,其中已发表但未被ISBT收录的2例,分别为ABO*B.01 with c.28+5885C>T、ABO*B.01 with c.28+5875C>T,另外3例为本研究首次报道的ABO*B新等位基因,分别为ABO*B.01(with c.3G>C)、ABO*B.01(with c.28+5862A>G)、ABO*B.01(with c.204-3C>G),均已被Genebank收录并公布。结论研究阐释了25例B亚型/AB亚型的分子生物学机制,并发现了5例ABO*B新等位基因。该研究丰富了ABO*B等位基因基因库,为更好地制定输血策略提供了理论依据。

Objective To study the genotype and molecular biological characteristics of 25 samples which showed B/AB subtypes.Methods The ABO*B alleles were identified by PCR-SSP or ABO exons 6-7/1-7 were analyzed by Sanger sequencing technology.Unidentified samples were further detected by PacBio:SMRT sequencing technology.Results Genetic sequence analysis of 25 B subtype/AB subtype samples revealed 7 known ABO*B alleles,namely ABO*BW.27(3/25,12.00%),ABO*BW.03(5/25,20.00%),ABO*B3.03(2/25,8.00%),ABO*BEL.03(1/25,4.00%),ABO*BW.07(7/25,28.00%),ABO*B3.05(1/25,4.00%),ABO*BW.12(1/25,4.00%).There were 5 unknown ABO*B alleles,of which 2 published but not included in ISBT were ABO*B.01 with c.28+5885C>T,ABO*B.01 with c.28+5875C>T.The other 3 were ABO*B novel alleles reported for the first time in this study,namely ABO*B.01(with c.3G>C)、ABO*B.01(with c.28+5862A>G)、ABO*B.01(with c.204-3C>G).Those have been compiled and published by Genebank.Conclusion The study clarified the molecular biological mechanism of 25 B/AB subtype cases and identified 5 novel ABO*B alleles.It enriches the ABO*B gene bank and provides a theoretical basis for better transfusion strategies.