Optimization and clinical validation of a Real-Time PCR protocol for direct detection of Trichomonas vaginalis in pooled urine samples

A new Real- Time PCR protocol for the detection of Trichomonas vaginalis in pooled urine samples has been optimized and validated. 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. Versluiset 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). 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%). 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.