ObjectiveTo explore the isolation and antimicrobial susceptibility of Mycoplasma from urogenital tract, and provide the basis for rational clinical treatment.MethodsUrogenital tract specimens from 57 904 outpatients in a hospital between 2008 and 2014 were performed Ureaplasma urealyticum (Uu) and Mycoplasma hominis(Mh) culture, identification, and antimicrobial susceptibility testing.ResultsOf 57 904 patients with urogenital infection，21 614 (37.33%) had positive culture for Mycoplasma, isolation rate of Mycoplasma in female and male were 42.14% （18 917/44 889）and 20.72% （2 697/13 015）respectively; Mycoplasma was mainly isolated from population of 21-40 years old; Uu≥104 CFU/mL and Mh<104 CFU/mL mixed infection was common(69.35%). The resistance of Mycoplasma to doxycycline, josamycin, and tetracycline were all low(<10%); resistance rates of Mh to doxycycline, erythromycin, clarithromycin, and roxithromycin were all significantly higher than Uu （all P<0.05）.ConclusionMycoplasma infection/carriage rate in female outpatients is significantly higher than male outpatients, antimicrobial profile of Uu is different from Mh, josamycin and doxycycline can be as the first choice for treatment of nongonococcal urethritis (cervicitis) caused by Mycoplasma.