Detection of the −1082 G/A Promoter Polymorphism of Interleukin-10 Gene by ARMS-PCR Technique among some Iraqi Rheumatoid Arthritis Patients

Abstract

Rheumatoid arthritis (RA) is a chronic inflammatory disease in which interleukin (IL)-10 gene plays an important role. There are, however, controversial reports that IL-10 promoter polymorphism may be an independent marker of susceptibility and severity of RA. The aim of the present study was to examine the IL-10 promoter polymorphism among RA patients. The aim of this study was to investigate the association between IL-10 G-1082A promoter polymorphism of interleukin-10 gene in some Iraqi rheumatoid arthritis patients using ARMS-PCR technique.We examined 70 RA patients (60 females and 10 males; aged: 20 – 74 years, mean: 51.7 years) were recruited from a private clinic in different region of Salahaddin governorate and 50 healthy individuals (40 females and 10 males; age: matching with patients) who were the staff of Tikrit University and hospital workers. The venous blood samples (5 ml) were collected from all the participants at the time of clinical examination, aliquoted in plain and EDTA vacutainers for the determination of erythrocyte sedimentation rate, C-reactive protein, presence of RFs. Genomic DNA was isolated from the whole-blood samples of all the patients and control subjects. The IL-10 -1082 G/A polymorphism genotyping Amplification Refractory Mutation System- Polymerase Chain Reaction (ARMS-PCR) method was used.ESR rate and RF positivity Levels of CRP were found as high as ≥ 20, 86.3% among patients, while all the healthy individuals were negative for this test except for a few number.The genotype GA for the −1082 G/A polymorphism of the IL-10 gene promoter was the most frequently observed genotype in both patients and controls, however, the distribution of the GG and AA genotype in patients and control was different. While genotype AA has a higher frequency than GG in patients (27.04vs.23.04), genotype GG was more frequently observed than AA in controls (46.08vs. 0.08).