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Journal of Clinical Microbiology, November 2001, p. 3819-3822, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3819-3822.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

TaqMan-Based Detection of Trichomonas vaginalis DNA from Female Genital Specimens

Jeanne A. Jordan,1,2,* Donna Lowery,1,3 and Massimo Trucco4

Magee-Women's Research Institute1 and Department of Pathology,2 Department of Obstetrics, Gynecology, and Reproductive Sciences,3 and Department of Pediatrics,4 University of Pittsburgh, Pittsburgh, Pennsylvania

Received 5 April 2001/Returned for modification 7 June 2001/Accepted 12 August 2001

A double-labeled fluorescent probe was designed and evaluated for detecting Trichomonas vaginalis DNA in a 5' nuclease (TaqMan) assay. The T. vaginalis-specific probe contains a 5'-fluorescein (5'-FAM) and a 3'-rhodamine (TAMRA) derivative. Female genital secretions were collected on Amplicor (Roche Molecular, Indianapolis, Ind.) swabs and by a transport system used for Chlamydia trachomatis and/or Neisseria gonorrhoeae DNA detection by PCR. Five hundred fifty-two female genital specimens, of which 248 (45%) were vaginal specimens and 304 (55%) were introital, were tested for both T. vaginalis DNA and viable microorganisms using the 5' nuclease assay and broth culture, respectively. Of these, 304 of 552 (55%) were also evaluated by direct microscopic examination for the characteristic motile organism. After resolving discrepancies, the comparisons produced an analytical sensitivity and specificity for the TaqMan-based PCR assay of 97.8 and 97.4%, respectively. As a result, Delta RQ values (differences in fluorescence due to probe hybridization and resulting 5'-FAM cleavage from the specific PCR product) of >= 2.0 and <= 1.5 were established for T. vaginalis-positive and -negative cutoffs, respectively. Delta RQ values between 1.5 and 2.0 were considered indeterminate. Overall findings revealed a high level of agreement between PCR and culture for detecting T. vaginalis. Potential benefits of the 5' nuclease assay include a greater sensitivity compared to direct microscopic examination and the ease of testing large numbers of clinical specimens in a significantly shorter turnaround time compared to culture.

* Corresponding author. Mailing address: Magee-Women's Research Institute, 204 Craft Ave., Pittsburgh, PA 15213. Phone: (412) 641-4104. Fax: (412) 641-6156. E-mail: jordanja+{at}pitt.edu.

Journal of Clinical Microbiology, November 2001, p. 3819-3822, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3819-3822.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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