Rapid genotyping of human platelet antigen 5 with fluorophore-labelled hybridization probes on the LightCycler.

Abstract

Genotyping of human platelet antigens (HPAs) can be useful for the diagnosis and therapy of alloimmune thrombocytopenic syndromes such as neonatal alloimmune thrombocytopenic purpura, post-transfusion purpura and refractoriness to platelet transfusion therapy. We developed a single-tube method for HPA-5 genotyping on the LightCycler that combines rapid-cycle polymerase chain reaction with allele-specific fluorescent probe melting for mutation detection. Our method is fast, robust and suitable for routine HPA-5 typing. This work extends recent studies on HPA-1 typing using the LightCycler.