Thirty-one middle ear fluids were obtained for Chlamydia trachomatis culture by tympanocentesis or myringotomy in 26 children and acute otitis media (AOM) or persistent otitis media with effusion (POME). The study population included 12 patients with AOM and 14 patients with POME. C. trachomatis was recovered from the middle ear fluid of two patients (16%) with AOM and from one patient (7%) with POME. Sixty external auditory canal specimens from 30 normal children were cultured. None was positive for C. trachomatis. Because our study population was small the true significance of this organism as a pathogen in ADM or POME is impossible to assess at this time.