BACKGROUND: RNA simultaneous amplification and testing (RNA-SAT) is a sensitive, specific, and rapid test method for the detection of pathogens in clinical samples. Herein, we used RNA-SAT to understand the detection status and distribution of common pathogens causing infections in patients with clinical urethritis to provide early clinical diagnosis and treatment. METHODS: Urine or swab samples were collected from 1,594 patients with suspected urinary tract infections between April 2022 and August 2023. RNA-based detection of common urogenital pathogens, Chlamydia trachomatis, Ureaplasma urealyticum, Neisseria gonorrhoeae, and Mycoplasma genitalium, was performed. Infection rates in different genders and ages were analyzed. Results of pathogen cultures were recorded during the same period. RESULTS: Majority of the patients included in the study were outpatients (99.25%, 1,582/1,594). Out of the 1,594 pa-tients, 828 (51.94%) had a urogenital pathogen infection. U. urealyticum had the highest detection rate (37.50%, 555/1,480), followed by C. trachomatis (17.13%, 223/1,302), N. gonorrhoeae (10.45%, 127/1,215), and M. genitalium (9.98%, 127/1,273). Coinfections of U. urealyticum, C. trachomatis, and/or N. gonorrhoeae were observed in 13.56% (174/1,283) of tested samples. Total RNA detection rate did not significantly differ between sexes
however, the detection rate of U. urealyticum and N. gonorrhoeae in females was significantly higher and lower than in males, respectively. Further, detection rate showed a decreasing trend with age. Finally, the RNA-SAT approach showed a higher detection rate for U. urealyticum, N. gonorrhoeae, and M. genitalium than the conventional culture approach did. CONCLUSIONS: U. urealyticum and C. trachomatis are the main pathogens responsible for urogenital tract inflammation. RNA-SAT can identify major pathogen infections at the early stages of the disease, effectively compensate for the deficiency of culture-based detection, and aid in early clinical diagnosis, and hence intervention.