To investigate the value of clinical application of simultaneous amplification and testing of RNA (SAT-RNA) for detecting (CT) and (UU) by comparing with the polymerase chain reaction testing of DNA (PCR-DNA) method. Specimens from both urethra swab and the first avoid urine which should be at least one hour after the previous urination were collected from 163 men who were scheduled for fertilization and embryo transfer (IVF-ET) treatment due to female factors at Center for Reproductive Medicine, Shengjing Hospital of China Medical University during the period of April 2016 to April 2017. Among the 163 men, 109 simultaneously provided semen that was collected after 3-7 days of sexual abstinence for the testing. Urine and semen specimens were detected for CT and UU with SAT-RNA, while urethra swab specimens were detected for CT and UU with standard PCR-DNA. Detection results of the SAT-RNA were compared with those of the PCR-DNA method. The positive rate of UU in the urethra swab detected with PCR-DNA and that