Aim: The aetiology of tympanosclerosis is not yet clear. This prospective, controlled, clinical study investigated the relationship between Helicobacter pylori and tympanosclerosis aetiology.Materials and methods: The study included 14 patients with tympanosclerosis and 26 with other forms of chronic otitis media. All patients underwent surgery for chronic otitis media. Mucosal biopsies were taken, and examined for H pylori using the Campylobacter-Like Organism (CLO) test.Results: Tympanoplasty was performed in 29 patients (72.5 per cent), radical mastoidectomy in eight (20 per cent) and myringoplasty in three (7.5 per cent). The presence of H pylori was tested in all tympanosclerosis biopsies, but in only 26.9 per cent of biopsies from other forms of chronic otitis media. A statistically significant difference in H pylori presence was found (p ≥ 0.01).Conclusion: This study represents a preliminary investigation of the association between H pylori and tympanosclerosis development. © 2011 JLO (1984) Limited.