Abstract: Objective To compare the detection rates of fluorescent-quantitative PCR, culture-drug susceptibility test and pure culture in detection of Mycoplasma, provide scientific evidence for the clinical diagnosis Mycoplasma infection. Methods The detection of Mycoplasma infection in cervix was conducted for 58 women with 3 tests mentioned above. Results The detection rate of fluorescent-quantitative PCR was 39.7% (23/58), the detection rate of culture-drug susceptibility test was 32.8% (19/58) and the detection rate of pure culture was 19.0% (11/58). The differences on the detection rates of 3 tests were statistical significant (P0.05). Conclusion Fluorescent-quantitative PCR has highest value in laboratory diagnosis, culture-drug susceptibility test has high value in clinical diagnosis. Although pure culture has less value in laboratory diagnosis compared with the first 2 tests, it still has practical value in medical institutions with limited technical conditions.