Optimisation of HLA-B27 Testing by Association of Flow Cytometry and DNA Typing |
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Authors: | G Bonnaud C Aupetit P M Preux M Cogné M Drouet |
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Institution: | (1) Laboratoire d’Immunologie et Immunogénétique, FR;(2) EP CNRS 118, Faculté de Médicine, and, FR;(3) Laboratoire de Biostatistiques, Faculté de Médecine, CHRU Dupuytren, Limoges, France, FR |
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Abstract: | HLA-B27 typing contributes to the diagnosis of ankylosing spondylitis. The classical technique of microlymphocytotoxicity
is costly and can give false-negative results. We have compared 304 samples using two relatively new methods – flow cytometry
and PCR-SSP – and evaluated their respective uses in routine analysis. Flow cytometric HLA-B27 testing was performed using
three monoclonal anti-B27 antibodies (HLA-ABC-m3, GS145.2 and FD705 clones). Cut-off values were established to differentiate HLA-B27-positive from HLA-B27-negative samples
with ROC curves. Although flow cytometric analysis with a reliable monoclonal antibody (mAb) is valuable for HLA screening,
none of the HLA-B27 flow cytometry protocols was sufficient on its own to ascertain the HLA phenotype in 100% of samples.
Two false negatives were observed with the FD705 mAb and the use of two different monoclonal antibodies did not increase the
accuracy of HLA-B27 typing. HLA-B27 typing using molecular biology is a reliable but costly technique. Therefore we suggest
that DNA typing could be used as a complementary technique and applied to samples whose HLA-B27 phenotype cannot be determined
by flow cytometry. The association of flow cytometry and DNA typing is, in our experience, an economical and reliable approach.
Received: 12 February 1998 / Accepted: 13 July 1998 |
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Keywords: | :Ankylosing spondylitis – Flow cytometry – HLA-B27 – HLA-B27 subtypes – Polymerase chain reactions |
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