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 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.

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