Objective: To study the prevalence and antimicrobial susceptibility of Ureaplasma urealyticum (Uu) and Mycoplasma hominis (Mh) in Changsha, and provide laboratory evidence for clinical drug use. Methods: A retrospective study was conducted to analyze 53 006 specimens of suspected genital mycoplasma infection in Xiangya Second Hospital of Changsha District and Hunan Wangwang Hospital from 2010 to 2017, and to analyze the infection rate and drug resistance rate of Uu and Mh. Results: From 2010 to 2017, a total of 53 006 specimens were detected, where there were 16 830 cases of Uu infection, the infection rate was 31.75%; 2 471 cases of Mh infection, the infection rate was 4.66%; and 1 071 cases of Uu and Mh mixed infection, the infection rate was 2.02%. Male Uu infection rate was 19.48%(5 989/30 749), which was lower than the female infection rate 48.71%(10 841/22 257) (χ²=5 091, P<0.001); male Mh infection rate was 3.16%(973/30 749), lower than female infection rate 6.73%(1 498/22 257) (χ²=369,P<0.001). The population of genital mycoplasma infection is concentrated between 20 and 40 years old, accounting for 71.76% (12 077/16 830). The drug resistance rates of Uu and Mh to doxycycline and minocycline were less than 2%, while the drug resistance rate to quinolones was higher; The resistance rate of Uu to macrolide antibiotics such as erythromycin, josamycin and clarithromycin were less than 2%, while the resistance rate to azithromycin, erythromycin and roxithromycin were higher, 29.74%(5 006/16 830) and 53.74%(9 045/16 830), respectively, and the resistance rate of Mh to macrolide antibiotics (except josamycin) was higher than 90%.Between 2010 and 2017, a gradually increasing resistance of ureaplasmas to azithromycin, from 3.81% (46/1 206) in 2010 to 53.15% (1 503/2 828) in 2017, and decreasing resistance to gatifloxacin and thiamphenicol were observed, from 76.78% (926/1 206) and 60.28% (727/1 206) in 2010 decreased to 34.23% (968/2 828) and 37.87% (1 071/2 828) in 2017, respectively. The resistance rate of Mh to gatifloxacin and thiamphenicol were decreased, from 68.93% (122/177) and 41.81% (74/177) in 2010 to 53.54% (159/297) and 21.21% (63/297) in 2017, respectively. Conclusions Doxycycline, minocyclinum and josamycin are good treatment options for genital mycoplasma in Changsha. The resistance rate of Uu to azithromycin is increasing, suggesting that the abuse of azithromycin is present in Changsha, and indicating that better management of antibiotics is necessary.