GENETIC TYPING OF HUMAN PLATELET ANTIGEN-1 (HPA-1) BY OLIGONUCLEOTIDELIGATION ASSAY IN A SPECIFIC AND RELIABLE SEMIAUTOMATED SYSTEM

Genotyping of platelet alloantigens with the possibility of using any type of cellular material as a source of DNA has become a preferred pr ocedure, particularly in thrombocytopenic patients when platelet count s are too low for phenotyping. Recently human platelet antigen 1 (HPA- 1) has been identified as an inherited risk factor for coronary thromb osis. The different detection methods currently used have disadvantage s for large-scale DNA diagnosis, including the need for electrophoresi s (allele-specific restriction enzyme analysis, amplification with seq uence-specific primers) or the potential risk of reduced specificity ( allele-specific oligonucleotide hybridization), In this report we desc ribe the adaptation of an automated oligonucleotide ligation assay to genotype HPA-1 in polymerase chain reaction (PCR)-amplified DNA sample s. HPA-1a and HPA-1b phenotypes corresponded to the results of the dif ferent genotyping assays. The genotypes determined with the ELISA-base d PCR-oligonucleotide ligation assay were in 100% concordance with the results obtained by conventional allele-specific restriction enzyme s ite analysis and PCR amplification with sequence-specific primers, The automated oligonucleotide ligation assay provides a rapid, reliable, nonisotopic method to genotype human platelet antigens that can rapidl y be applied to large population screening.