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Journal of Clinical Microbiology, December 2003, p. 5619-5622, Vol. 41, No. 12
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.12.5619-5622.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Use of the Roche LightCycler Instrument in a Real-Time PCR for Trichomonas vaginalis in Urine Samples from Females and Males

Justin Hardick,1 Samuel Yang,2 Shin Lin,2 Della Duncan,1,3 and Charlotte Gaydos1*

Division of Infectious Disease,1 Department of Emergency Medicine, Johns Hopkins University School of Medicine,2 Baltimore City Health Department, Baltimore, Maryland3

Received 25 June 2003/ Returned for modification 26 August 2003/ Accepted 21 September 2003

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.

* Corresponding author. Mailing address: 1159 Ross Bldg., 720 Rutland Ave., Baltimore, MD 21205. Phone: (410) 614-0932. Fax: (410) 614-9775. E-mail: cgaydos{at}jhmi.edu.

Journal of Clinical Microbiology, December 2003, p. 5619-5622, Vol. 41, No. 12
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.12.5619-5622.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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