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Clinical Chemistry 41: 687-692, 1995;
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Clinical Chemistry, Vol 41, 687-692, Copyright © 1995 by American Association for Clinical Chemistry

Validation of allele-specific polymerase chain reaction for DNA typing of HLA-B27

HM Steffens-Nakken, G Zwart and FA van den Bergh
Department of Clinical Chemistry, Hospital Medisch Spectrum Twente, Enschede, The Netherlands.

To find a specific method for HLA-B27 typing for the diagnosis of rheumatic disorders, we extensively tested the single-step B27-specific polymerase chain reaction (PCR) described by Dominguez et al. (Immunogenetics 1992;36:277-82). This method, which relies on specific primer recognition of a sequence in the third exon (unique to the B27- allele), was used for screening of 270 characterized blood samples, 57 of which were B27-positive. The method proved to be both sensitive and specific: It unambiguously identified all B27-positive samples and produced no false-positive results. For approximately 1% of the samples, we had to repeat DNA isolation and PCR to obtain a clear control amplification signal. In contrast to the specificity of the PCR method, parallel-performed flow cytometry gave ambiguous results in 3% of the samples because of antibody cross-reactivity. Flow cytometry and the PCR method described were similar in labor and costs. Therefore, we conclude that the proposed single-step PCR is feasible in a routine laboratory and would improve the reliability of HLA-B27 typing.


The following articles in journals at HighWire Press have cited this article:


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X Zhu and Z Yang
TNF{alpha} gene variants and ankylosing spondylitis
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X. Zhu, Y. Wang, L. Sun, Y. Song, F. Sun, L. Tang, Z. Huo, J. Li, and Z. Yang
A novel gene variation of TNF{alpha} associated with ankylosing spondylitis: a reconfirmed study
Ann Rheum Dis, November 1, 2007; 66(11): 1419 - 1422.
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M. A.M. Bon, A. van Oeveren-Dybicz, and F. A.J.T.M van den Bergh
Genotyping of HLA-B27 by Real-Time PCR without Hybridization Probes
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J. J. M. L. Hoffmann and W. C. M. Janssen
HLA-B27 phenotyping with flow cytometry: further improvement by multiple monoclonal antibodies
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