Objectives: To evaluate diagnostic value of high frequency single prop tone tympanometry (1000 Hz) in otitis media with effusion in infants. Method: We selected 90 infants patients aged 4 to 8 months with long standing or recurrent attacks of otitis media with effusion at the outpatient clinic of ENT Department, Aljumhory teaching hospital, Mosul, Iraq, May 2018 to June 2019. Immittancemetry included single low probe tone (226Hz) and high probe tone (1000Hz) frequency tympanometry at same setting were conducted. Tympanometric pattern occurrence, acoustic reflex thresholds and peak compensated static acoustic admittance for each tested frequency were estimated and analyzed. Result: At 226 Hz frequency 25 patients were single peak tympanogram and 65 patients were flat curve, while at 1000 Hz frequency 51 patients were single peak and 39 patients had flat curve. Acoustic reflex threshold were present in 45 patients and absent in the other 45 at 226 Hz frequency, but it was present in 60 infants and absent in 30 infants at the 1000 Hz. Mean static peak acoustic admittance at 226 Hz was 0.69, while at 1000 Hz the mean was 4.52. Peak not founded in 31 patients at 226 Hz & in 19 patients at 1000 Hz. Conclusion: 1000 Hz probe tone tympanometry was more efficient in the diagnosis of otitis media with effusion.