HPA genotyping by PCR-SSP: report of 4 exercises.
OBJECTIVE
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.


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%.


CONCLUSIONS
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.
Metcalfe
P
P
National Institute for Biological Standards and Control, Potters Bar, UK. pmetcalfe@nibsc.ac.uk
Cavanagh
G
G
Hurd
C
C
Ouwehand
W H
WH
D016824:Antigens, Human Platelet / Q000235:genetics*; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*
D016824:Antigens, Human Platelet