To determine frequencies of HLA DRβ1 alleles in rheumatoid arthritis in Pakistani patients. Cross sectional/analytical study. Department of Immunology, Armed Forces Institute of Pathology, Rawalpindi in collaboration with Rheumatology departments of Military Hospital, Rawalpindi and Fauji Foundation Hospital, Rawalpindi, from January 2009 to January 2010. HLA DRβ1 genotyping of one hundred Pakistani patients, diagnosed as having RA as per American College of Rheumatology revised criteria 1987, was done. HLA DRβ1 genotyping was carried out at allele group level (DRβ1*01-DRβ1*16) by sequence specific primers in RA patients. Comparison of HLA DRβ1 allele frequencies between patients and control groups was made using Pearson's chi-square test to find possible association of HLA DRβ1 alleles with RA in Pakistani rheumatoid patients. HLA DRβ1*04 was expressed with significantly increased frequency in patients with rheumatoid arthritis (p <0.05). HLA DRβ1*11 was expressed statistically significantly more in control group as compared to rheumatoid patients indicating a possible protective effect. There was no statistically significant difference observed in frequencies of HLA DRβ1 allele *01, DRβ1 allele *03, DRβ1 allele *07, DRβ1 allele *08, DRβ1 allele *09, DRβ1 allele*10, DRβ1 allele *12, DRβ1 allele *13, DRβ1 allele *14, DRβ1 allele *15 and DRβ1 allele *16 between patients and control groups. The identification of susceptible HLA DRβ1 alleles in Pakistani RA patients may help physicians to make early decisions regarding initiation of early intensive therapy with disease modifying anti rheumatic medicines and biological agents decreasing disability in RA patients.