HLA-B*27 subtyping by PCR-RFLP in Spanish patients with ankylosing spondyiitis |
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Authors: | A. Fraile,J. Martin,M.A. Lö pez-Nevot,L. Mataran,A. Nieto |
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Affiliation: | Instituto de Parasitologia y Biomedicina "Lopez Neyra". CSIC, Granada, Spain.;Hospital Virgen de las Nieves, Granada, Spain |
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Abstract: | Abstract: We describe the use of restriction analysis on PCR-amplified DNA for detecting all B*27 subtypes except B*2710 and B*2711 (i.e. from B*2701 to B*2709). After detecting B*27 by Sty I, double digestions consisting of Sty I plus another informative enzyme led to subtype assignment. We used mismatched primers to create restriction sites when necessary. The method avoids group-specific amplifications and other laborious optimization procedures. It was successfully tested on a panel of well characterized cell lines covering different B*27 subtypes. Then, we studied a group of 57 ankylosing spondyiitis patients and 746 controls from the south of Spain. B*27 showed a very strong association with the disease (OR=211.27, P=0˜7). B*2702 and B*2705 distribution in controls (20% and 77.1%, respectively) differed from previously reported data in the Spanish population. We unexpectedly found the B*2707 allele in our population (one control). |
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Keywords: | ankylosing spondyiitis HLA-B*27 subtypes PCR-RFLP |
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