Genotyping of the Human Platelet Antigen‐1 by ELISA Detection of Allele – Specific Amplicons

Publisher Website
Google Scholar
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.