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 of 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 previously described have required different reaction mixes and PCR conditions. We have designed a set of sequence-specific primers for HPA 1-5 which react optimally under identical reaction and PCR conditions. Comparative testing with reference samples gave 100% concordance. The advantages of this method include speed; accuracy; smaller sample requirements and no reliance on human typing sera or platelet integrity. The method also has the potential to be applied to amniotic fluid. Simplified DNA techniques will lead to more extensive and proficient platelet antigen typing. This will facilitate accurate laboratory diagnosis of alloimmune thrombocytopenia and the provision of HPA-matched blood products.
Cavanagh
G
G
National Blood Service, Newcastle, Newcastle Upon Tyne, UK.
Dunn
A N
AN
Chapman
C E
CE
Metcalfe
P
P
D000483:Alleles; D016824:Antigens, Human Platelet / Q000032:analysis / Q000235:genetics*; D004247:DNA / Q000032:analysis* / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods
D016824:Antigens, Human Platelet; D004247:DNA