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  • Title: Genotyping of the human platelet antigen-1 by ELISA detection of allele-specific amplicons.
    Author: Müller TH, Döscher A, Schunter F.
    Journal: Vox Sang; 1997; 73(3):185-8. PubMed ID: 9358622.
    Abstract:
    BACKGROUND AND OBJECTIVES: 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. MATERIALS AND 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. CONCLUSION: This method offers efficient HPA-1 genotyping in large numbers of donors and patients.
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