UNIT 6.22 Flow Cytometric Screening for the HLA-B27 Antigen on Peripheral Blood Lymphocytes

  1. Wilfried H.B.M. Levering1,
  2. Kees Sintnicolaas1,
  3. Henk Wind2,
  4. Herbert Hooijkaas2,
  5. Jan W. Gratama2

Published Online: 1 AUG 2005

DOI: 10.1002/0471142956.cy0622s33

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Levering, W. H., Sintnicolaas, K., Wind, H., Hooijkaas, H. and Gratama, J. W. 2005. Flow Cytometric Screening for the HLA-B27 Antigen on Peripheral Blood Lymphocytes. Current Protocols in Cytometry. 33:6.22:6.22.1–6.22.12.

Author Information

  1. 1

    Laboratory for Histocompatibility and Immunogenetics, Sanquin Blood, Bank South West Region, Rotterdam, The Netherlands

  2. 2

    Erasmus MC, Rotterdam, The Netherlands

Publication History

  1. Published Online: 1 AUG 2005
  2. Published Print: JUL 2005


HLA-B27 is common to the entire group of seronegative spondyloarthropathies, and assessment of HLA-B27 is of diagnostic importance if any of these diseases are considered. Among the alternatives to traditional typing by the complement-dependent cytotoxicity assay, flow cytometric HLA-B27 screening is most widely used in general diagnostic laboratories, as it is simple, rapid, and cost effective. This unit describes screening for HLA-B27 on peripheral blood lymphocytes using more than one HLA-B27 monoclonal antibody to detect possible cross-reactivity with non-HLA-B27 antigens. Screening is hampered by the lack of true monospecific anti-HLA-B27 monoclonal antibodies. Cross-reactivities of anti-HLA-B27 with other HLA-B antigens have been reported. The authors recommend use of GS145.2 and FD705 antibodies, as this combination avoids most false-positive conclusions. Lyse-and-wash sample processing is employed. A multiparameter flow methodology is applied. Three to four parameters–forward scatter, side scatter, HLA-B27 expression, and, in some cases CD3 or B7 expression–are acquired.


  • HLA-B27;
  • flow cytometry;
  • cross-reactivity;
  • HLA-typing;
  • monoclonal antibody