Development and validation of a genotyping kit for the eight principal human platelet alloantigen systems

Thrombocytopenia in newborns is often due to maternal alloimmunization agai nst platelet alloantigens of the foetus which have been inherited from the father and are absent in the mother. Our aim was to develop a "ready-to-use " typing kit based on polymerase chain reactions, using sequence-specific p rimers for rapid and simultaneous genotyping of the eight principal human p latelet alloantigens. The typing technique uses two specific primer pairs f or each bi-allelic system and a monomorphic primer pair as amplification co ntrol, with a single temperature cycle programme and identical PCR stringen cy conditions for all pairs of primers. This kit allows typing of blood sam ples of small volume within three hours after reception. Validation criteri a are essential to check the reliability and specificity of the test, and D NA controls carrying the targeted HPA alleles must be obtained from typed i ndividuals or created in vitro by PCR. (C) 2000 Editions scientifiques et m edicales Elsevier SAS.