HLA‐B*27 typing by PCR—restriction fragment length polymorphism

In this study we describe the use of PCR-RFLP for genotyping HLA-B*27. A 557 bp fragment from HLA-B locus is amplified and subjected to digestion with StyI. The presence of B*27 is detected on electrophoresis by the appearance of 431 + 126 bp pattern. The same pattern could be obtained only for the very infrequent allele B*7301. However, this allele was not amplified in the B73 sample tested with the primers and conditions used in this study. Nevertheless, we have designed two PCR-RFLP approaches for separating these alleles. The PCR-RFLP method was tested on a panel of forty-three cell lines and applied to fifty spondyloarthritic patients and one-hundred-eighty healthy subjects. Given its robustness, technical simplicity and cost-effectiveness, we think that this method can be incorporated for routine use in most laboratories.