A(1,2)BO(1,2) GENOTYPING BY MULTIPLEXED ALLELE-SPECIFIC PCR

The ABO blood group is the most clinically important human alloantigen system in transfusion medicine. The system involves three antigens A, B and H. H antigen is converted to either A or B by the activity of a lpha 1 --> 3-N-acetyl-galactosaminyl transferase (A transferase) or al pha 1 --> 3 galactosyl transferase (B transferase). The O phenotype is the result of an inactive glycosyltransferase, which is unable to gly cosylate the H antigen. The immunological properties of the ABO system were identified at the turn of the century; however, the genetic basi s of the ABO system has only recently been characterized. This has ena bled the development of a number of molecular ABO typing methods. Desc ribed here is a two-reaction multiplex allele-specific PCR (ASPCR) gen otyping assay for the A(1), A(2), B, O-1 and O-2 subtypes. 11 differen t allele-specific oligonucleotide primers were selected to detect the presence or absence of the O-1 associated G --> (-) deletion at base 2 61, the O-2 associated G --> A substitution at base 802, the B associa ted G --> A substitution at base 803, and finally the A(2) associated C --> (-) deletion at base 1059. A total of 122 peripheral blood sampl es were genotyped and serologically forward and reverse typed. A conco rdance rate of 98.4% (120/122 samples) was observed between the actual genotype and the serologically-based predicted genotype. These result s indicate that this assay provides a rapid, accurate, and simple meth od for A(1,2)BO(1,2) genotyping that serves as a useful supplement to standard serological ABO typing.