Objective To investigate the infection of genital tract derived Mycoplasma and antibiotic resistance in local area infertility patients. Methods The mycoplasm from genital tract in patients who had visited and tested in sterility clinic from the year 2011 to 2013 were cultured and identified. The antibiotic resistance was also studied. Results There was 410 positive cases in the 848 samples, and 370 patients were infection by Ureaplasma urealytium (Uu). Uu infection rates in male and female were 45.6% and 24.1%, respectively. The drug resistance rate of ciprofloxacin and ofloxacin were 89.0% and 73.3%, respectively. The drugs with high susceptibility were Doxycycline, Pristinamycin, Tetracycline and Josamycin, the susceptibility rate of these drugs reached more than 90%. Conclusion The genital tract Uu infection rate of infertility patients and the resistance rate of Ciprofloxacin and Ofloxacin are very high in local area. There was a significant difference of infection rates in male and female. The clinic treatment should be based on the drug sensitivity test results.