Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 1 3 2009 09 15 12 15 EN WB van Leeuwen Erasmus Medical Center Rotterdam Department of Medical Microbiology & Infectious Diseases‘s- Gravendijkwal 2303015 CE Rotterdam, The Netherlands. WHA Zandijk Erasmus Medical Center Rotterdam Department of Medical Microbiology & Infectious Diseases‘s- Gravendijkwal 2303015 CE Rotterdam, The Netherlands. 2015 10 01 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. https://ijm.tums.ac.ir/index.php/ijm/article/view/23 https://ijm.tums.ac.ir/index.php/ijm/article/download/23/23