HLA-B27 is an antigen associated with the disease ankylosing spondylit
is. Ninety percent of Caucasians with ankylosing spondylitis possess t
he HLA-B27 antigen. However, only 20% of Caucasians with the HLA-B27 a
ntigen will develop the disease. Defining the presence or absence of t
he HLA-B27 antigen can be helpful in differentiating ankylosing spondy
litis from juvenile rheumatoid arthritis, In this study, we evaluated
the application of two monoclonal antibodies (MoAbs), using flow cytom
etric analysis for the detection of HLA-B27 antigen. After an initial
comparison of HLA-B27 analysis by flow cytometry to the standard micro
lymphocytotoxicity assay, cutoffs were established to differentiate HL
A-B27 positive from HLA-B27 negative. One MoAb showed very reliable re
sults with >99% accuracy in discriminating HLA-B27 positive from HLA-B
27 negative samples, Various parameters were investigated to obtain th
e optimum results and showed that incubation time, reagent lot, and th
e flow cytometric instrument can affect the results, We concluded that
a reliable MoAb and flow cytometry are valuable for the rapid and ine
xpensive determination of HLA-B27 typing in the clinical setting, Howe
ver, testing conditions can affect the accuracy of results; therefore,
adequate parallel testing in various conditions must be performed in
order to establish the proper standards. (C) 1995 Wiley-Liss, Inc.