Background: Many A and B suballeles responsible for ABO subgroup formation have been identified. Some of these minor alleles have mutations in the ABO gene coding sequence. Most of these mutations are due to single-nucleotide substitution and lead to amino acid alteration. Several alleles at the ABO locus appear to be caused by crossing over between dissimilar alleles.
Study design and methods: Blood samples were collected from an individual with AelBel phenotype and her family members. Sequencing of the seven ABO exons was performed on these samples. The following was performed for the samples from the AelBel proposita: cloning and sequencing of the genomic DNA of the ABO gene, reverse transcription-polymerase chain reaction (PCR) analysis of cDNA transcript of the ABO gene, sequence-specific priming (SSP)-PCR analysis and direct sequencing of the ABO gene, SSP-PCR DNA typing of generic HLA-ABC and HLA-DRB, and short-tandem repeat (STR)-PCR typing on 15 autosomal, 2 X-chromosomal, and 6 Y-chromosomal loci.
Results: The proposita with AelBel phenotype has blood group chimerism with a major group of A1v-O1v/O1(O01) and a minor group of B(B101)/O1v(O02). Additional haplotypes on HLA-ABC, HLA-DR-B, STR loci, and Y-chromosome STR loci were present on the proposita. The paternal genotype is B(B101)/O1(O01) and the maternal genotype is A1v(A102)/O1v(O02). No other siblings have the A1v-O1v hybrid allele. Parentage was confirmed with the paternity STR-PCR test. Full-length cDNA transcripts of the B(B101) allele and alternately spliced cDNA transcripts of the hybrid A1v-O1v, O(1), and O1v alleles were cloned from the proposita. The A1v-O1v hybrid gene contains two missense mutations: C467T and G829A, resulting in Pro156Leu and Val277Met substitution.
Conclusion: Formation of the A1v-O1v hybrid allele appears to result from de novo recombination in the germ line of the mother during meiosis. G829A with Val277Met appears to be responsible for the decrease in A-transferase activity and Ael phenotypic expression in the proposita. The chimeric minor population of B(B101)/O1v is responsible for Bel phenotypic expression in the proposita.