HLA-B*27 subtyping by PCR-RFLP in Spanish patients with ankylosing spondylitis.
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 spondylitis patients and 746 controls from the south of Spain. B*27 showed a very strong association with the disease (OR=211.27, P=10(-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).
Fraile
A
A
Instituto de Parasitología y Biomedicina Lopez Neyra, CSIC, Granada, Spain.
Martin
J
J
López-Nevot
M A
MA
Mataran
L
L
Nieto
A
A
D015796:HLA-B27 Antigen / Q000145:classification / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013030:Spain; D013167:Spondylitis, Ankylosing / Q000235:genetics* / Q000276:immunology
D015796:HLA-B27 Antigen