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HLA-B27 determination using serological methods. A comparison of enzyme immunoassay and a microlymphocytotoxic test with flow cytometry and a molecular biological assay

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Abstract

Typing for HLA-B27 is routinely performed in patients with seronegative spondarthritides. Besides the microlymphocytotoxic test (MLCT), other serological techniques have been developed such as enzyme immunoassays (EIA) using serum or plasma as a source for the determination of soluble HLA-B27 (sHLA-B27) and flow cytometric (FC) methods. The aim of the present study was to check the accuracy and reliability of the EIA for sHLA-B27 in comparison to the MLCT using antibodies against HLA-B27 and cross-reacting specificities (CRS), as well as an FC method and a molecular biological method. Any discrepant results should be typed with the MLCT using a complete panel of anti-HLA-class I antibodies, with FC and with a molecular biological technique. The EIA should also be repeated in those patients, using serum and plasma from a new venipuncture. In 81 patients with rheumatic disorders, the EIA and the MLCT using antibodies against HLA-B27 and CRS were performed. Based on the MLCT with a complete panel of anti-HLA-class I antibodies as a standard, discrepant test results were obtained for 9 out of 81 patients with the MLCT using antibodies against HLA-B27 and CRS and with the EIA. The following wrong results occurred: in the MLCT with anti-HLA-B27 and CRS, there were two false-negative results; in the EIA there were four false-negative and one false-positive results: one sample was undeterminable. In comparison with the MLCT, including the complete panel of HLA-class I antibodies, as well as with a molecular biological technique, typing with FC showed a complete concordance. Our investigations demonstrated that for routine typing for HLA-B27 the MLCT cannot be replaced by EIA because of a significant number of mistypings. The MLCT performed only with antibodies against HLA-B27 and CRS may also lead to typing errors. No errors were detected using flow cytometry. If only serological methods can be performed in a laboratory a combination of flow cytometry and MLCT could therefore enhance the safety of HLA-B27 typing.

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Authors and Affiliations

  1. 5th Department of Internal Medicine, Wilhelminenspital, Vienna

    A. Dunky & E. Wagner

  2. Ludwig Boltzmann Institute for Rheumatology and Balneology, Kurbadstrasse 10, P.O. Box 78, A-1107, Vienna-Oberlaa, Austria

    J. Neumüller

  3. Central Laboratory, Wilhelminenspital, Vienna, Austria

    C. Hübner & P. M. Bayer

  4. Clinical Department for Blood Group Serology, University of Vienna, Vienna, Austria

    G. F. Fischer, D. W. M. Schwartz & W. R. Mayr

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  1. A. Dunky
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  2. J. Neumüller
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  3. C. Hübner
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  4. G. F. Fischer
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  6. E. Wagner
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  7. D. W. M. Schwartz
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  8. W. R. Mayr
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Dunky, A., Neumüller, J., Hübner, C. et al. HLA-B27 determination using serological methods. A comparison of enzyme immunoassay and a microlymphocytotoxic test with flow cytometry and a molecular biological assay. Rheumatol Int 16, 95–100 (1996). https://doi.org/10.1007/BF01409980

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  • DOI: https://doi.org/10.1007/BF01409980

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