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  • Title: Flow cytometry immunophenotyping and polymerase chain reaction-site-specific primers genotyping for HPA-1 alloantigens in an Italian blood donor population.
    Author: Tazzari PL, Cirillo D, Bontadini A, Ricci F, Masi R, Conte R.
    Journal: Vox Sang; 1998; 74(1):42-5. PubMed ID: 9481860.
    Abstract:
    BACKGROUND AND OBJECTIVES: 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. MATERIALS AND 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.
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