ABO GLYCOSYLTRANSFERASE GENOTYPING BY POLYMERASE CHAIN-REACTION USINGSEQUENCE-SPECIFIC PRIMERS

Serological typing for the classical ABO blood groups is routinely per formed using anti-A and anti-B antisera of polyclonal or monoclonal or igin, which are able to distinguish four phenotypes (A, B, AB, and O). Modern molecular biology methods offer the possibility of direct ABO genotyping without the need for family investigations. Typing can be d one with small amounts of DNA and without detection of blood group mol ecules on the surface of red blood cells. We developed a system of eig ht polymerase chain reactions (PCR) to detect specific nucleotide sequ ence differences between the ABO alleles O-1, O-2, A(1) A(2), and B. P CR amplification using sequence-specific primers and detection of ampl ification products by agarose gel electrophoresis is one of the fastes t genotyping methods and is easy to handle. With our method we tested the A(1,2)BO(1,2) genotypes of 300 randomly chosen persons out of a po ol of platelet donors and found the results to be consistent with ABO glycosyltransferase phenotypes. We also identified a presumably new AB O allele, which may be the result of a crossing-over event between all eles O-1 and A(2). (C) 1996 by The American Society of Hematology.