HPA genotyping by PCR-SSP: Report of 4 exercises

Background and Objectives: Polymerase chain reaction with sequence specific primers (PCR-SSP) is widely used for the determination of the alleles enco ding the human platelet antigens (HPA)-1 to 5. In order to evaluate and imp rove performance with this technique, four exercises were organised during 1996-1998. Materials and Methods: Coded DNA samples were distributed from t he National Institute for Biological Standards and Control (NIBSC) as follo ws: exercise one, 18 samples; two, 12 samples; three, 6 samples, and four, 4 samples. Results: Performance improved over the four exercises following the adoption of a consensus protocol and the re-design of one primer. The p ercentage of incorrect results in each exercise was as follows: exercise on e, 9%; two 3.2%, three, 0.8%, and four, 0.3%. Conclusion: The modified PCR- SSP protocol is a reliable method for genotyping HPA-1 to 5 in reference la boratories. DNA-based HPA genotyping has an important role in platelet immu nology and further exercises will be included in the bi-annual platelet imm unology exercises organised by NIBSC.