title: Genotyping of the human platelet antigen systems 1 through 5 by multiplex polymerase chain reaction and ligation-based typing.
abstract: BACKGROUND
  Platelet-specific antibodies may be involved in refractoriness to platelet transfusions, disorders such as neonatal alloimmune thrombocytopenia, and post-transfusion purpura. Genotyping for the major human platelet antigen (HPA) systems HPA-1 through HPA-5 is of considerable help in establishing the diagnoses of these diseases.
  
  
  METHODS
  A new genotyping method is described. Alleles of all five systems are amplified in a multiplex polymerase chain reaction. Subsequently, aliquots of the amplification products are thermocycled in the presence of a pair of allele-specific oligonucleotide probes and a heat-stable ligase. After heat denaturation, the probes hybridize adjacent to complementary sequences of the amplification product. In a perfect match, the two probes become covalently joined. Detection of the ligation product is performed with an enzyme-linked immunosorbent assay.
  
  
  RESULTS
  Complete concordance of genotypes between the ligation-based typing and established genotyping methods was determined in 54 Austrian (HPA-1, -2, -3, and -5) and 56 Japanese (HPA-4) individuals. Ligation-based genotyping of HPA-1 polymorphism using platelet-derived RNA as starting material gave concordant results in all 15 cases tested.
  
  
  CONCLUSIONS
  Multiplex polymerase chain reaction in combination with ligation-based typing allows fast typing of large numbers of platelet donors and screening for critical antigens in pregnant women.
authors.affiliation: Department of Transfusion Medicine, University of Göttingen, Germany.
authors.forename: T J
authors.initials: TJ
authors.lastname: Legler
authors.forename: M
authors.initials: M
authors.lastname: Köhler
authors.forename: W R
authors.initials: WR
authors.lastname: Mayr
authors.forename: S
authors.initials: S
authors.lastname: Panzer
authors.forename: H
authors.initials: H
authors.lastname: Ohto
authors.forename: G F
authors.initials: GF
authors.lastname: Fischer
mesh_terms: D001483:Base Sequence; D001792:Blood Platelets / Q000276:immunology*; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000235:genetics*; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000235:genetics*; D016133:Polymerase Chain Reaction
chemical_list: D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D019038:Platelet Glycoprotein GPIb-IX Complex