Development of a PCR method to detect HLA-B27 in ankylosing spondylitis

Abstract: The aim of the project was to develop a PCR method to detect HLA-B27 at the Immunology Department of St. James hospital in Dublin. The HLA-B27 gene is common among patients with ankylosing spondylitis (AS). Ninety percent of patients with AS have the HLA-B27 gene and it is therefore counted as a risk factor and could be used as part of the diagnosis. Twenty-two frozen blood samples from patients with AS or suspected AS were donated from the rheumatology department at St. James hospital. PCR is a well known and common technique, many hospital laboratories have a PCR machine and therefore PCR is a good choice for detection of the HLA-B27 gene. A multiplex PCR was developed where a PCR control, primers to the β-globin gene, was used in the same tube as the HLA-B27 primers, to secure that the PCR worked in every tube. Finally a blind test was performed to test the specificity of the PCR. The result shows that the specificity was 100%. Of all patient samples, sixteen was HLA-B27 positive and six were HLA-B27 negative. In addition, optimal conditions for the PCR and the way to extract DNA from frozen blood were successfully established. For future diagnosis, the described PCR can be used to detect the HLA-B27 gene in patients and it can be considered as a start for further development of a real-time PCR for detection of the HLA-B27.