title: Genotyping of the human platelet antigen-1 by ELISA detection of allele-specific amplicons.
abstract: OBJECTIVE
  The purpose of the study was to establish a valid and efficient method for genotyping of the human platelet alloantigen-1 (HPA-1) in large sample numbers.
  
  
  METHODS
  Digoxigenin- and fluorescein-labelled allele-specific primers and a biotinylated common primer were included in the hot-start PCR. Amplicons were bound to avidin-coated plates to identify the products by ELISA.
  
  
  RESULTS
  This approach reduced the number of PCR analyses for HPA-1 typing by half. PCR products were detected with high sensitivity and good reproducibility (interassay-CV: < 8%) in the ELISA. The typing of 100 blood donors with both PCR plus gel electrophoresis and ELISA-PCR showed identical results.
  
  
  CONCLUSIONS
  This method offers efficient HPA-1 genotyping in large numbers of donors and patients.
authors.affiliation: German Red Cross Blood Transfusion Service, Institute Oldenburg, Germany. ckmthm@nord.de
authors.forename: T H
authors.initials: TH
authors.lastname: Müller
authors.forename: A
authors.initials: A
authors.lastname: Döscher
authors.forename: F
authors.initials: F
authors.lastname: Schunter
mesh_terms: D000483:Alleles; D016824:Antigens, Human Platelet / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D016824:Antigens, Human Platelet