Background/aim: Acute rheumatic fever and rheumatic heart disease are major causes of morbidity and mortality in developing countries. Genetic studies have determined that the immune response in rheumatic heart disease is genetically controlled and that there is a close relationship between the gene of concern and the class II human leukocyte antigen (HLA) gene. The aim of this study was to evaluate the relationship of serum HLA-B alleles and tumor necrosis factor alpha (TNF-∝) with rheumatic heart disease. Materials and methods: A total of 50 consecutive patients with rheumatic heart disease and 50 controls were enrolled in the study. HLA alleles were analyzed using sequence-specific primer-polymerase chain reaction and nucleotide sequencing. Results: The HLA-B35 allele was significantly more common in patients with rheumatic heart disease than the control group (P = 0.043). The HLA-B44 allele was significantly more common in control patients than in patients with rheumatic heart disease (P = 0.014). There was a significant inverse correlation between high-sensitivity C-reactive protein and mitral valve area (P = 0.001). There was no correlation between TNF-∝ levels and mitral valve area (P = 0.066). Conclusion: Our findings confirmed the association between HLA-B alleles and rheumatic heart disease.