Aim. To determine the frequency of detection of conjunctival C. trachomatis (CT), M. hominis (MH), and U. urealyticum (UU) infections in young adults with dry eye disease (DED), since these infections may potentially produce the chronic subclinical inflammation characteristic of DED. Materials and Methods. The study included subjects of 25-45 years of age, divided into the DED (n = 114) and nondry eye control (n = 98) groups, with the diagnosis based on self-reported complaints, biomicroscopy, the Schirmer I test, and break-up time. All patients had conjunctival scrapings taken to detect CT, MH, and UU with direct fluorescent-antibody assay kits. Results. At least one of the three microorganisms was found in 87.7% of the DED patients versus 8.2% of the controls. Of all the DED patients, 63.2%, 50.8%, and 42.1% were found to be infected with CT, MH, and UU, respectively. Multiple pathogens were identified in 65% of the DED patients found to be infected. CT infection was detected in 6.1% of the controls. Conclusion. C. trachomatis, M. hominis, and U. urealyticum were detected with high frequency in the conjunctiva of young adults with DED and may be an important risk factor for DED in them.