Objective To investigate the infection of Mycoplasma in female urogenital system and the evolution of drug resistance. Methods Ureaplasma urealyticum (UU) and Mycoplasma hominis (MH) infections were examined by pathogen culture and fluorescence quantitative PCR in urogenital samples from 530 female patients in 2007, and drug sensitivity tests were performed in vitro. The results were compared with those in 2000. Results In 2000, there were 99 patients with Mycoplasma infections, in which 65 were of UU infections, 15 were of MH infections and 19 were of UU + MH infections. In 2007, there were 344 patients with Mycoplasma infections, in which 236 were of UU infections, 47 were of MH infections and 61 were of UU + MH infections. Patients aged 21～30 had the highest Mycoplasma infection rates both in 2000 (46.2%) and 2007 ( 50.5% ) ; while Mycoplasma infection rate in patients aged under 20 rose from 5.1% in 2000 to 12.8% in 2007 (χ2 = 4.682, P < 0.05). Both in 2000 and 2007, pathogens presented the highest drug resistance rates to tetracycline (TET) and erythromycin ( ERY ) which were all bore 80%. Compared with 2000, drug resistance rates to levofloxacin (LEV), azitromvcin (AZI) and ofloxacin (OFL) rose in 2007, and the differences were of statistical significance (P <0.05), while drug resistance rates to dmoxycycline (DOX), minocyclin (MIN) and josamycin (JOS) were still in the low level. Conclusions UU is the primary pathogen of infection in female urogenital system, and there is a tendency of Mycoplasma infection in younger age women. DOX, MIN and JOS can be the preferred medicines for Mycoplasma infections.