title: Use of the Roche LightCycler instrument in a real-time PCR for Trichomonas vaginalis in urine samples from females and males.
abstract: Trichomonas vaginalis is the agent of a highly prevalent sexually transmitted infection (STI) that can result in vaginitis, urethritis, and preterm birth. Traditional methods of diagnosis, including wet preparation, can be unreliable. In this study, we describe the adaptation of an existing PCR method for specific detection of T. vaginalis DNA into a rapid real-time PCR assay based on fluorescence resonance energy transfer (FRET) probe chemistry. The FRET-based assay described demonstrated high sensitivity with a detection limit of 1.06 organisms, as well as a high specificity. A total of 253 urine samples collected prospectively from both men and women were tested for T. vaginalis DNA with both the FRET-based assay and a previously validated PCR assay. When the validated PCR assay was used as the "gold standard" and after discrepancies had been resolved, our FRET-based assay demonstrated an analytical sensitivity and specificity of 90.1 and 100%, respectively. Overall results suggest that FRET-based assays can provide rapid, accurate, and high-throughput detection of T. vaginalis and may prove useful in clinical settings and for large-scale screening programs.
authors.affiliation: Division of Infectious Disease, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Justin
authors.initials: J
authors.lastname: Hardick
authors.forename: Samuel
authors.initials: S
authors.lastname: Yang
authors.forename: Shin
authors.initials: S
authors.lastname: Lin
authors.forename: Della
authors.initials: D
authors.lastname: Duncan
authors.forename: Charlotte
authors.initials: C
authors.lastname: Gaydos
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D014245:Trichomonas Infections / Q000175:diagnosis* / Q000652:urine; D014246:Trichomonas vaginalis / Q000235:genetics / Q000302:isolation & purification*; D014556:Urine / Q000469:parasitology*
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan