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Genotyping of the Human Platelet Antigen‐1 by ELISA Detection of Allele – Specific Amplicons
Author(s) -
Müller Thomas H.,
Döscher Andrea,
Schunter Friedrich
Publication year - 1997
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1046/j.1423-0410.1997.7330185.x
Subject(s) - genotyping , amplicon , typing , biotinylation , digoxigenin , microbiology and biotechnology , polymerase chain reaction , primer (cosmetics) , allele , biology , genotype , chemistry , genetics , gene , in situ hybridization , gene expression , organic chemistry
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.