Objective.To compare a real-time polymerase chain reaction (PCR) assay with broth culture for the detection ofTrichomonas vaginalisusing self-collected vaginal swabs.Methods.Self-collected vaginal swabs were obtained from adolescent and young adult African-American women participating in HIV-1 prevention programs.T. vaginalisculture was performed using the InPouch TV System. Samples for the real-time PCR assay were collected using the BDProbeTec ET Culturette Direct Dry Swab system and tested in a laboratory-developed assay which targeted a repeated sequence of the genome. Discrepant samples that were culture negative and positive in the real-time PCR assay were tested in a confirmatory PCR which targeted a different region of theT. vaginalisgenome, the18S ribosomal DNA gene.Results.Of the 524 specimens tested by both culture and real-time PCR, 36 were culture positive and 54 were positive in the real-time PCR assay; 16 of the 18 discrepant specimens were also positive in the confirmatory PCR assay. Using a modified gold standard of positive by culture or positive in both PCR assays, the sensitivity of the real-time PCR assay was 100% and the specificity was 99.6%, whereas culture had a sensitivity of 69.2% and a specificity of 100%.Conclusions.The real-time PCR assay was sensitive and specific for the detection ofT. vaginalisDNA from self-collected vaginal swab specimens. The ability to use the BDProbeTec dry swab system for the real-time PCR testing allowed for the detection ofChlamydia trachomatis, Neisseria gonorrhoeae,andT. vaginalisfrom a single specimen.