optimization and clinical validation of a real-time pcr protocol for direct detection of trichomonas vaginalis in pooled urine samples

background and objectives: a new real- time pcr protocol for the detection of trichomonas vaginalis in pooled urine samples has been optimized and validated. materials and methods: the amplification protocol, targeting a 2kb repeated gene in the t. vaginalis genome, was optimized by varying pcr parameters. as a reference method, a real-time pcr protocol targeting the beta-tubulin gene (y. versluis et al, 2006, int j std aids 17:642) was used. clinical validation was performed with pooled urine samples obtained from patients of the sexually transmitted diseases clinic of a university hospital (n=963; from february – june 2007). results: positive samples with the new optimized technique is 1.1%  (n=10), while the beta-tubulin real-time pcr method generated four positives (0.3%). conclusion: the new rt- pcr protocol is a sensitive (1.000) and specific (0.993) procedure to detect and to identify t. vaginalis in urine samples.