Objectives Our main objective was to determine the prevalence of bacterial etiologies of Otitis Media with Effusion (OME) among Omani children as well as to evaluate the antibiotic susceptibility to penicillins. Methods A prospective cross-sectional study was conducted on 28 Omani children under 5 years of age with OME. The Middle Ear Effusion (MEE) was sent for direct culture and polymerase chain reaction (PCR). Results Out of 38 MEE, 8(21.1%) showed growth on direct culture. The majority of bacteria were sensitive to penicillin. 17 (44.7%) samples were positive using PCR with 5 (29.4%) of them had more than one positive pathogen. The detection rates for H. Influenzae, M. catarrhalis and S. pnemoniae were 31.6%, 18.4% and 10.5% respectively. Conclusion The prevalence of bacteria causing otitis media with effusion among Omani children was not different than the rates reported worldwide however the order of the most common ones differs slightly. Keywords: Otitis media with effusion; Bacteria; Polymerase Chain Reaction.