RAPID DETECTION OF HLA-B27 SEQUENCE SPECIFIC ALLELES BY IN-HOUSE OPTIMIZED MOLECULAR ASSAY FOR THE DETECTION OF ANKYLOSING SPONDYLITIS

Abstract

Introduction: Ankylosing Spondylitis (AS) is a long term disease, involved in inflammation of the joints between the spinal bones and the joints between the spine and pelvis. Chronic inflammation in these areas causes pain and stiffness. Over the time, chronic inflammation of the spine can lead to a complete cementing together of the vertebrae, a process referred to as ankylosis.

Materials and Method: 40 blood specimens were collected from suspected patients with symptoms related to AS attending orthopedics and neurology departments of Shri Mahant Indiresh Hospital, Dehradun. Samples were processed using In house protocol by Conventional PCR using silica columns for high yield nucleic acid extraction. Results: Out of 40 specimens, 21came positive and remaining 19 were negative by both Olerup SSP approach and in house PCR protocol for HLA B-27 SSA. Age groups wise, positivity rate was maximum i.e. 66.6 % in the patients above 60 years of age. However, in the age group of 0–20 years, 21-40 years and 41–60 years, the positivity rate was 50, 52 and 62.5 %, respectively, which were comparatively less from the age group above 60 years. Discussion: In developing nations, the patients can’t afford for expensive molecular diagnostic tests. The In-House protocol standardized for the amplification of HLA-B27 is a novel, rapid, accurate and highly sensitive & specific technique for low cost as compared to commercially available assays. By using optimized protocol the cost can be reduced to 2-3 folds and hence will help in early and accurate diagnosis and management of the disease.

Keywords: Sequence Specific Primers, Ankylosing Spondylitis, Seronegative Spondyloarthropathy,  Internal Control, Amplicons.