HLA-B-ASTERISK-27 TYPING BY PCR-RESTRICTION FRAGMENT LENGTH POLYMORPHISM

In this study we describe the use of PCR-RFLP for genotyping HLA-B27. A 557 bp fragment from HLA-B locus is amplified and subjected to dige stion with Styl. The presence of B27 is detected on electrophoresis b y the appearance of 431+126 bp pattern. The same pattern could be obta ined only for the very infrequent allele B7301. However, this allele was not amplified in the B73 sample tested with the primers and condit ions used in this study. Nevertheless, we have designed two PCR-RFLP a pproaches for separating these alleles. The PCR RFLP method was tested on a panel of forty-three cell lines and applied to fifty spondyloart hritic patients and one-hundred-eighty healthy subjects. Given its rob ustness, technical simplicity and cost-effectiveness, we think that th is method can be incorporated for routine use in most laboratories.