Trichomoniasis is a significant sexually transmitted disease (STD) in the spectrum of public health and primary care because of its association with agents such as human immunodeficiency virus and Neisseria gonorrhoeae. However, its true significance may be underestimated due to diagnostic modalities that exhibit poor sensitivity. A total of 1,086 genital specimens from two urban emergency departments, a suburban urgent-care facility, and a metropolitan outpatient physician group were subjected to transcription-mediated amplification-based Trichomonas vaginalis analyte-specific-reagent (ASR) testing (Gen-Probe, Inc.). The rate of positive molecular ASR results (14.5%) doubled that of direct saline preparation (7.0%; P < 0.0002). Analogous increases were observed at one emergency department and within the outpatient physician group (P < 0.0002). No significant increase in the rate of positive molecular ASR results was observed from the facilities that encountered a lower frequency of black/African American patients. While positive T. vaginalis findings via direct saline preparation did not have a significant association with concomitant Chlamydia trachomatis or N. gonorrhoeae infection overall, a positive T. vaginalis ASR result was a better predictor of concomitant C. trachomatis or N. gonorrhoeae infection (odds ratios of 2.34 and 4.46, respectively; P < 0.0001). The increased rate of positive T. vaginalis ASR results was observed in both point-of-care (P ؍ 0.02 versus direct saline preparation) and laboratory (P ؍ 0.003) testing. Highly sensitive T. vaginalis molecular ASR not only transcends issues of specimen integrity and microscopic acumen but also has an increased ability to predict the likelihood of additional STDs in defined populations.In spite of the discovery of Trichomonas vaginalis nearly 175 years ago and documentation of its inhabitation of the female urogenital tract and the male urinary tract in the late 1800s, pathogenicity was not ascribed to this agent until the European literature of the 20th century (19). Reports have since shown the significance of antecedent T. vaginalis infection, especially in human immunodeficiency virus coinfection (22, 37), acquisition (21), and transmission (17, 23); in pregnancy-related complications (10, 43); and in associations with pelvic inflammatory disease (16) and Neisseria gonorrhoeae infection (16,24). T. vaginalis is currently thought to be responsible for approximately 50% of all curable infections worldwide (5); worldwide estimates of annual trichomoniasis incidence have reached 180 million cases (42).While the aforementioned data may be of tremendous significance, trichomoniasis prevalence rates, both worldwide and in the United States, are thought to be grossly underestimated. Schwebke and Burgess (32) hypothesize that the variable sensitivity of T. vaginalis diagnostic testing contributes partially to these artificially low statistics. Direct examination of genital saline collections continues to serve as a common basis for laboratory detection...