The identification of sexually transmitted pathogens by rapid and sensitive techniques is of interest due to the spread routes including the risks of acquisition and transmission. Our aim was to develop multiplex real-time PCR assays to simultaneously detect M. hominis, M. genitalium, U. urealyticum, N. gonorrhoeae, and C. trachomatis. Clinical samples from female patients with presumptive diagnosis of infection with these bacteria were examined. Two multiplex real-time PCRs were developed: MI: N. gonorrhoeae, C. trachomatis, U. urealyticum; MII: M. hominis, M. genitalium, U. urealyticum. The reactions were able to detect at least one of the agents, especially three or more microrganisms in a single clinical sample describing co-infections. The detection limits were 29.7 copies/L for N. gonorrhoeae, 30.1 copies/L for U. urealyticum, 29.9 copies/L for C. trachomatis, 29.7 copies/L for M. hominis and 30.4 copies/L for M. genitalium. The multiplex tests developed in this study multiplex real-time PCRs provided a novel qualitative method to detect simultaneously sexually transmitted pathogens. Although additional studies with a greater number of clinical samples are needed, the results were notably encouraging, and these methods can be used as valuable tool in routine clinical laboratories.
