HPA GENOTYPING BY PCR SEQUENCE-SPECIFIC PRIMING (PCR-SSP) - A STREAMLINED METHOD FOR RAPID ROUTINE INVESTIGATIONS

We describe a streamlined method for the simultaneous identification o f alleles of the human platelet antigens (HPA) 1-5. The method employs the polymerase chain reaction with sequence specific primers (PCR-SSP ). Although PCR-SSP has been applied to HPA genotyping, all methods pr eviously described have required different reaction mixes and PCR cond itions. We have designed a set of sequence-specific primers for HPA 1- 5 which react optimally under identical reaction and PCR conditions. C omparative testing with reference samples gave 100% concordance. The a dvantages of this method include speed; accuracy; smaller sample requi rements and no reliance on human typing sera or platelet integrity. Th e method also has the potential to be applied to amniotic fluid. Simpl ified DNA techniques will lead to more extensive and proficient platel et antigen typing. This will facilitate accurate laboratory diagnosis of alloimmune thrombocytopenia and the provision of HPA-matched blood products.