ABO glycosyltransferase genotyping by polymerase chain reaction using sequence-specific primers

• Published in: Blood 1996-09, Vol.88 (5), p.1852-1856
• Main Authors: GASSNER, C, SCHMARDA, A, NUSSBAUMER, W, SCHÖNITZER, D
• Format: Article
• Language: English
• Subjects: ABO Blood-Group System - genetics; Alleles; Base Sequence; Biological and medical sciences; Blood Grouping and Crossmatching - methods; DNA Primers; Electrophoresis, Agar Gel; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Genotype; Humans; Immunohematology; Molecular Sequence Data; N-Acetylgalactosaminyltransferases - genetics; Polymerase Chain Reaction; Red blood cell immunology
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.v88.5.1852.1852

Serological typing for the classical ABO blood groups is routinely performed using anti-A and anti-B antisera of polyclonal or monoclonal origin, 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 done with small amounts of DNA and without detection of blood group molecules on the surface of red blood cells. We developed a system of eight polymerase chain reactions (PCR) to detect specific nucleotide sequence differences between the ABO alleles O1, O2, A1, A2, and B. PCR amplification using sequence-specific primers and detection of amplification products by agarose gel electrophoresis is one of the fastest genotyping methods and is easy to handle. With our method we tested the A1,2BO1,2 genotypes of 300 randomly chosen persons out of a pool of platelet donors and found the results to be consistent with ABO glycosyltransferase phenotypes. We also identified a presumably new ABO allele, which may be the result of a crossing-over event between alleles O1 and A2.