Genetic Typing of Human Platelet Antigen 1 (HPA-1) by Oligonucleotide Ligation Assay in a Specific and Reliable Semi-Automated System

Br J Haematol. 1997 Jan;96(1):198-203. doi: 10.1046/j.1365-2141.1997.d01-1980.x.

Abstract

Genotyping of platelet alloantigens with the possibility of using any type of cellular material as a source of DNA has become a preferred procedure, particularly in thrombocytopenic patients when platelet counts are too low for phenotyping. Recently human platelet antigen 1 (HPA-1) has been identified as an inherited risk factor for coronary thrombosis. The different detection methods currently used have disadvantages for large-scale DNA diagnosis, including the need for electrophoresis (allele-specific restriction enzyme analysis, amplification with sequence-specific primers) or the potential risk of reduced specificity (allele-specific oligonucleotide hybridization). In this report we describe the adaptation of an automated oligonucleotide ligation assay to genotype HPA-1 in polymerase chain reaction (PCR)-amplified DNA samples. HPA-1a and HPA-1b phenotypes corresponded to the results of the different genotyping assays. The genotypes determined with the ELISA-based PCR-oligonucleotide ligation assay were in 100% concordance with the results obtained by conventional allele-specific restriction enzyme site 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 rapidly be applied to large population screening.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Human Platelet / genetics*
  • Enzyme-Linked Immunosorbent Assay
  • Genotype*
  • Humans
  • Oligonucleotides / metabolism*
  • Polymerase Chain Reaction
  • Restriction Mapping

Substances

  • Antigens, Human Platelet
  • Oligonucleotides