Genotyping of the human platelet antigen-1 by ELISA detection of allele-specific amplicons.
OBJECTIVE
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.


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.


CONCLUSIONS
This method offers efficient HPA-1 genotyping in large numbers of donors and patients.
Müller
T H
TH
German Red Cross Blood Transfusion Service, Institute Oldenburg, Germany. ckmthm@nord.de
Döscher
A
A
Schunter
F
F
D000483:Alleles; D016824:Antigens, Human Platelet / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D016824:Antigens, Human Platelet