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

ABSTRACT

Thrombocytopenia in newborns is often due to maternal alloimmunization against 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 primers for rapid and simultaneous genotyping of the eight principal human platelet alloantigens. The typing technique uses two specific primer pairs for each bi-allelic system and a monomorphic primer pair as amplification control, with a single temperature cycle programme and identical PCR stringency conditions for all pairs of primers. This kit allows typing of blood samples of small volume within three hours after reception. Validation criteria are essential to check the reliability and specificity of the test, and DNA controls carrying the targeted HPA alleles must be obtained from typed individuals or created in vitro by PCR.