An optimized method for routine HLA-B27 screening using flow cytometry

Autor: J. Albrecht, E M Veys, P. Lancaster, A. Falkenrodt, L. Buchner, L. Noens, N. Mir, F. de Keyser, R. Becker, F. Hulstaert, C. Costello, J. Kunz, I. Hannet, M. Tongio, Kenneth Strauss
Rok vydání: 1994
Předmět:
Zdroj: Cytometry. 18:21-29
ISSN: 1097-0320
0196-4763
DOI: 10.1002/cyto.990180106
Popis: Flow cytometry and monoclonal antibodies are promising tools for HLA-antigen detection. Previous approaches have been hampered by the lack of a carefully standardized system for calibration and sample analysis. A new system for HLA-B27 screening was developed using a FACScan flow cytometer, software for automated calibration and analysis, calibration beads, and the anti-HLA-B27-FITC/anti-Leu4-PE (CD3) monoclonal antibodies. The median fluorescence channel result for the HLA-B27-FITC signal of CD3+ T lymphocytes is compared to a decision marker. Values lower than this threshold are read as HLA-B27 negative and those above are recommended for retesting with the classic microcytotoxicity assay on the presumption of HLA-B27 positivity. The anti-HLA-B27 antibody reacts with all six HLA-B27 subtypes and shows a weaker binding to HLA-B7. The screening test results were compared with those from the microcytotoxicity assay for HLA-typing in studies involving several European centers. The observed sensitivity was 100% (95% Cl:98.6-100) and the specificity was 97.4% (95% Cl: 96.4-98.3). Other performance studies verified the reproducibility and reliability of results obtained with the screening system.
Databáze: OpenAIRE