Platelet-specific antibodies are involved in refractoriness to platelet transfusions, neonatal alloimmune thrombocytopenia, and posttransfusion purpura. Binding to different epitopes on platelet glycoproteins, human platelet antigens (HPA), they can cause severe thrombocytopenia. The polymorphic DNA regions of the best known HPA systems HPA-1 to HPA-5 were amplified from genomic DNA in a multiplex polymerase chain reaction (PCR). An automatable ligation-based assay (LCR) using allele-specific oligonucleotide probes was developed for rapid genotyping. The results were correlated with the well established allele-specific restriction enzyme assay (ASRA), and the phenotype was determined by the monoclonal antibody-specific immobilization of platelet antigens (MAIPA). We found a 100% concordance of genotypes between the HPA-LCR in 54 cases of NAIT, PTP and normal controls with the established methods. The gene frequencies calculated from 216 German and 55 Japanese random blood donors were similar to those determined by other time-consuming typing methods in a Caucasian and Japanese population, respectively. Multiplex PCR and the allele-specific HPA ligation assay provide a reliable tool for typing both small series and all platelet donors of a transfusion center in order to identify rare genotypes necessary for the treatment of patients suffering from NAIT, PTP or refractoriness to platelet transfusions.

Keywords:
HPA genotyping, Multiplex PCR, Ligation-based typing
This content is only available via PDF.
© 1995 S. Karger AG, Basel
1995
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.