4.2 Article

The detection of the HLA-B27 antigen by immunomagnetic separation and enzyme-linked immunosorbent assay - comparison with a flow cytometric procedure

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JOURNAL OF IMMUNOLOGICAL METHODS
卷 255, 期 1-2, 页码 15-22

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ELSEVIER SCIENCE BV
DOI: 10.1016/S0022-1759(01)00414-8

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ankylosing spondylitis; HLA-B27 antigen; flowcytometry assay; IMS-ELISA; polymerase chain reaction

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The HLA-B27 antigen is an important genetic marker in ankylosing spondylitis (AS). Methods for the detection of B27 include the microlymphocytotoxicity test and, more recently, flowcytometry (FC). Here, we describe a new method, IMS-ELISA, for measuring the B27-antigen. It combines immunomagnetic separation (IMS), to obtain B27-positive cells from whole blood samples, with an enzyme-linked immunosorbent assay (ELISA) as a read-out. IMS-ELISA was tested on 367 samples obtained from five different hospitals in Taiwan. The sensitivity, specificity and accuracy of the method were compared with FC. Any conflicting data between IMS-ELISA and FC was confirmed by HLA-DNA typing via PCR-SSP (polymerase chain re action-sequence specific primers). Overall, the results for sensitivity, specificity and accuracy obtained by IMS-ELISA and FC did not show any significant difference (p > 0.05). However, when considering laboratory time, cost, ease of operation and the screening of large samples for HLA-B27, the IMS-ELISA was superior to the FC method. We conclude that IMS-ELISA may be used as a fast screening method for HLA B27 detection. (C) 2001 Elsevier Science B.V. All rights reserved.

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