A rapid and simple ABO genotype screening method using a novel B/O2 versus A/O2 discriminating nucleotide substitution at the ABO locus.

An ABO genotype screening method discriminating the common alleles A1, A2, B, O1 and O2 at the ABO locus was made possible by the discovery of a novel nucleotide substitution (G1096A) present only in B and O2 alleles. A rapid and reliable single-tube approach using multiplex PCR with four primers amplifying exons 6 and 7 of the ABO genes followed by simultaneous addition of two restriction enzymes was developed and validated in a population of 150 Swedish blood donors. This technique is the most cost-efficient and informative ABO genotyping method reported to date.