HPA genotyping by PCR-SSP: report of 4 exercises

Abstract

Background and objectives: Polymerase chain reaction with sequence specific primers (PCR-SSP) is widely used for the determination of the alleles encoding the human platelet antigens (HPA)-1 to 5. In order to evaluate and improve performance with this technique, four exercises were organised during 1996-1998.

Materials and methods: Coded DNA samples were distributed from the National Institute for Biological Standards and Control (NIBSC) as follows: 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 percentage of incorrect results in each exercise was as follows: exercise one, 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 laboratories. DNA-based HPA genotyping has an important role in platelet immunology and further exercises will be included in the bi-annual platelet immunology exercises organised by NIBSC.