Molecular genetic analysis and identification of novel alleles of ABO subtypes / 中国输血杂志

ABSTRACT

【Objective】 To study the molecular mechanism of 95 samples of serological ABO subtypes. 【Methods】 A total of 95 samples with discrepancy between forward and reverse blood grouping were subjected to serological confirmation, and genotyped by polymerase chain reaction with sequence-specific primers (PCR-SSP). For those subtype alleles could not be detected by PCR-SSP, ABO gene exon 1-7 sequencing and gene single strand sequencing were performed successively to determine the mutation site and the gene location. 【Results】 A total of 34 ABO alleles were detected in 95 samples. Five common ABO alleles (ABO*A1.01, ABO*A1.02, ABO*B.01, ABO*O.01.01 and ABO*O.01.02) and 29 rare ABO alleles were identified, including 16 named alleles by ISBT (ABO*A2.01, ABO*A2.05, ABO*A2.13, ABO*A3.07, ABO*AW.37, ABO*AEL.05, ABO*B3.01, ABO*B3.05, ABO*BW.03, ABO*BW.07, ABO*BW.27, ABO*BEL.03, ABO*cisAB.01, ABO*cisAB.05, ABO*BA.02, ABO*BA.04) and 5 named alleles by dbRBC(A223, B309, Bw37, Bel09, Bw40)and eight unnamed alleles [ABO*B.01+ 978C>A, ABO*A1.02+ 248A>T, ABO*B.01+ 125dupT, ABO*B.01+ (98+ 1G>A), ABO*A1.02/ABO*B.01+ 1A>G, ABO*A1.02/ABO*O.01.01+ 28G>T, ABO*A1.02/ABO*B.01+ 538C>T, ABO*A1.02/ABO*O.01.01+ 797insT] .The last four samples could not be verified by single strand because of insufficient samples. In 95 samples, 76 samples (21 named alleles of ISBT and dbRBC) were identified by PCR-SSP, and the remaining 19 samples were identified by exon 1-7 sequencing of ABO gene, of which 8 were identified as unnamed alleles, and the remaining 11 samples were not identified as subtype alleles. 【Conclusion】 The molecular genetic mechanism of 95 serological ABO subtypes was revealed, and 8 rare novel alleles were identified. The detection of ambiguous blood groups is influenced by factors such as patient pathology and physiology, therefore the combination of serological testing and genetic testing is suggested for the identification of ABO subtype.