title: Flow cytometry immunophenotyping and polymerase chain reaction-site-specific primers genotyping for HPA-1 alloantigens in an Italian blood donor population.
abstract: OBJECTIVE
  There is increasing interest in the development of rapid and reliable techniques for human platelet alloantigen (HPA) typing. This study investigates the reliability of flow cytometry for large-scale immunophenotyping of platelet alloantigens.
  
  
  METHODS
  We used flow cytometry and polymerase chain reaction-site-specific primer (PCR-SSP) for the characterization of the human platelet antigen 1 (HPA-1) mosaic in blood donors.
  
  
  RESULTS
  By using specific alloantisera and immunofluorescence labelling 9 (2.6%) out of 351 samples were HPA-1a-negative. To confirm this antigenic phenotype, all of the latter samples were submitted to PCR-SSP analysis, showing an HPA1-b/b genomic pattern. In HPA-1a-positive donors, flow cytometry was unable to distinguish HPA-1a/b heterozygous from HPA-1a/a homozygous subjects who were clearly identified by genotyping.
  
  
  CONCLUSIONS
  Flow cytometry is a valuable tool for large-scale screening to identify HPA-1a-negative persons, whereas genotyping is the assay of choice for zygosity testing, antenatal diagnosis, and for thrombocytopenic alloimmunized patients.
authors.affiliation: Servizio di Immunoematologia e Trasfusionale, Policlinico S. Orsola-Malpighi, Bologna, Italy.
authors.forename: P L
authors.initials: PL
authors.lastname: Tazzari
authors.forename: D
authors.initials: D
authors.lastname: Cirillo
authors.forename: A
authors.initials: A
authors.lastname: Bontadini
authors.forename: F
authors.initials: F
authors.lastname: Ricci
authors.forename: R
authors.initials: R
authors.lastname: Masi
authors.forename: R
authors.initials: R
authors.lastname: Conte
mesh_terms: D000328:Adult; D016824:Antigens, Human Platelet / Q000097:blood* / Q000235:genetics / Q000276:immunology; D001782:Blood Donors; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D005838:Genotype; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D016130:Immunophenotyping; D039661:Integrin beta3; D007558:Italy; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D016824:Antigens, Human Platelet; C061248:ITGB3 protein, human; D039661:Integrin beta3