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Hla-B27 Typing Using Simple Allele Specific PCR in Suspectedspondyloarthritis Patients Visiting Diff

Anil Kumar Sah 1 , Pragya Hatawal* 2 , Sandip Shrestha* 2 , Laxmi Sharma 1 , Geeta Shrestha Vaidya 2 , Sunil Poudel, Paveen Nepal, Basant Pant

1 Annapurna Research Center, Maitighar, Kathmandu

2 SAAN int’l College, Gairidhra, Kathmandu

3 Annapurna Neurological Institutes and Allied Sciences, Kathmandu

7 Kathmandu Center for Genomics and Research Laboratory, Kathmandu

*Equal Contribution

**Corresponding Author:-



Spondyloarthritis is a group of interrelated rheumatic diseases which mainly includes Reactive arthritis (ReA), Enteropathic spondylitis, Psoriatic Arthropathy (PsA), undifferentiated spondyloarthritis (uSpA), juvenile spondyloarthritis and Ankylosing spondylitis, (AS)- the prototype of spondyloarthiritis. It is a class of seronegative arthritis. Inflammatory in back pain is the main clinical manifestation of SpA and is characterized by dull, low back or buttock pain lasting longer than three months improving with exercise but not with rest, morning stiffness lasting longer than thirty minutes, and insidious onset. There is a marked variation of association between SpA and HLA B27 surface antigen. This association encouraged in testing of HLA B27 antigen as one of the diagnostic tools in this inflammatory disease.

Objectives:- The main objective of this study is to perform HLA-B27 typing using simple allele specific PCR in suspected spondyloarthritis patients visiting the different hospitals of Kathmandu.

Methods:- The whole blood samples of the suspected patients were collected from different hospitals and DNA was extracted. The test was performed using sequence specific primer (SSP)-PCR which is far more accurate than conventional serological approach like MLCT and FC tests.

Results:- From total of 89 sample subjected for Spondyloarthritis, 24 (27%) were HLA-B27 positive with the

frequency 4.2%, 20.8%, 37.5%, 12.5% and 25% in the <15 years, 16-25 years, 26-35 years, 36-45 years and >45 years age group respectively. Among the B27 positive patients, 16 were males and 8 were females. This study reported 0% prevalence in the HLA-B27 test for healthy subjects. The findings of our proposed study showed a low frequency of HLA B27 among suspected patients with higher prevalence in

male. This study showed that the testing of HLA-B27 allele in diagnosis of SpA diseases is of clinical usefulness.

Conclusion:- To conclude, the study found low frequency of HLA-B27 with higher prevalence in male among suspected patients visiting different hospitals of Kathmandu

Key words: Spondyloarthritis( SpA), Ankylosing spondylitis (AS), Seronegative, HLA-B27, SSP PCR, Microlymphocytotoxicity (MLCT), Flowcytometry (FC)

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