Chorioamnionitis or intra-amniotic infection is an infection that affects the intrauterine content during pregnancy. Numerous studies have reported vaginal colonization with various types of infectious agents as a risk factor for chorioamnionitis. Although this complication occurs due to the ascending polymicrobial bacterial infection at the time of membrane breakage, it may also occur in pregnant women with intact membranes, mainly due to Ureaplasma urealyticum (U. urealyticum) and Mycoplasma hominis (M. hominis). The main aim of the present study was to identify a region-specific panel of infectious agents that can be used more accurately determine premature birth, as well as the premature rupture of membranes (PROM). Thus, a 10-year retrospective study was conducted. A total of 1,301 pregnant women with PROM and premature birth or spontaneous abortion were included in the study. It was observed that the main infectious agent varied in the five groups analyzed in total. The infectious agent distribution also varied depending on environmental parameters. Ureaplasma was found to be the most frequently detected germ amongst the infectious agents of the vaginal cultures from pregnant women enrolled in the present study, regardless of gestational age. On the whole, the findings of the present study suggest that additional studies are required, in order to confirm that diagnosis and treatment according to laboratory results of vaginal infections with U. urealyticum/M. hominis during the first trimester of pregnancy could prevent premature birth, abortion or chorioamnionitis.