DNA typing of human platelet antigen systems 1, 2, 3 and 5 in B-lymphoblastoid cell lines of the International Histocompatibility Workshop

Alloimmunization against human platelet antigens (HPA) can cause thrombocytopenia in different clinical settings, including neonatal alloimmune thrombocytopenia, post transfusion purpura, and refractoriness to platelet transfusion. Recently, DNA based methods have been described for analysis of the polymorphism of the human platelet antigens. However, until now, no reference material for standardization purposes is available. We thus determined the DNA polymorphism of the human platelet antigen (HPA) systems 1, 2, 3 and 5 in B-lymphoblastoid cell lines collected for the International Histocompatibility Workshops. DNA typing of the HPA systems was done by PCR amplification with sequence-specific primers (PCR-SSP). A new enzyme (AmpliTaq Gold) was introduced to enhance the specificity of the PCR. We present a panel of five B-lymphoblastoid cell lines, representing all heterozygous and homozygous genotypes of the tested HPA systems. This work thus provides the reference material required for DNA based diagnosis of human platelet antigens.