Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1992 Feb;30(2):465–472. doi: 10.1128/jcm.30.2.465-472.1992

Development of a polymerase chain reaction-based diagnosis of Trichomonas vaginalis.

D E Riley 1, M C Roberts 1, T Takayama 1, J N Krieger 1
PMCID: PMC265079  PMID: 1537918

Abstract

We developed a polymerase chain reaction (PCR)-based test for detecting the protozoan parasite Trichomonas vaginalis. Genomic libraries were constructed from two independent clinical isolates of T. vaginalis. From these libraries, 12 genomic clones were purified, sequenced, and then screened for uniqueness by computer-assisted sequence comparisons. PCR reactions were performed to evaluate eight PCR-primer pairs, including a primer pair that targeted the T. vaginalis ferredoxin gene. All eight primer pairs yielded PCR products of the expected sizes. However, six of the primer pairs amplified their respective target sequences in limited numbers of clinical T. vaginalis isolates, suggesting the presence of significant genomic variability among isolates. An exception was a primer pair, termed TVA5-TVA6, that amplified a 102-bp genomic sequence, termed A6p, in all of 24 clinical isolates. The A6p sequence was not detected by PCR in human DNA or in a wide variety of flagellates, ciliates, or bacteria tested. The A6p sequence appears highly selective for a broad range of T. vaginalis isolates and holds promise for PCR-based diagnosis of the parasite.

Full text

PDF
472

Images in this article

468Image
on p.468
468Image
on p.468
469Image
on p.469
469Image
on p.469

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alderete J. F., Kasmala L., Metcalfe E., Garza G. E. Phenotypic variation and diversity among Trichomonas vaginalis isolates and correlation of phenotype with trichomonal virulence determinants. Infect Immun. 1986 Aug;53(2):285–293. doi: 10.1128/iai.53.2.285-293.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alderete J. F., Suprun-Brown L., Kasmala L. Monoclonal antibody to a major surface glycoprotein immunogen differentiates isolates and subpopulations of Trichomonas vaginalis. Infect Immun. 1986 Apr;52(1):70–75. doi: 10.1128/iai.52.1.70-75.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bauer H. M., Ting Y., Greer C. E., Chambers J. C., Tashiro C. J., Chimera J., Reingold A., Manos M. M. Genital human papillomavirus infection in female university students as determined by a PCR-based method. JAMA. 1991 Jan 23;265(4):472–477. [PubMed] [Google Scholar]
  4. Blanchard A. Ureaplasma urealyticum urease genes; use of a UGA tryptophan codon. Mol Microbiol. 1990 Apr;4(4):669–676. doi: 10.1111/j.1365-2958.1990.tb00636.x. [DOI] [PubMed] [Google Scholar]
  5. Cao M., Xiao X., Egbert B., Darragh T. M., Yen T. S. Rapid detection of cutaneous herpes simplex virus infection with the polymerase chain reaction. J Invest Dermatol. 1989 Mar;92(3):391–392. doi: 10.1111/1523-1747.ep12277232. [DOI] [PubMed] [Google Scholar]
  6. Fleury F. J. Diagnosis of Trichomonas vaginalis infection. JAMA. 1979 Dec 7;242(23):2556–2557. [PubMed] [Google Scholar]
  7. Fouts A. C., Kraus S. J. Trichomonas vaginalis: reevaluation of its clinical presentation and laboratory diagnosis. J Infect Dis. 1980 Feb;141(2):137–143. doi: 10.1093/infdis/141.2.137. [DOI] [PubMed] [Google Scholar]
  8. Genesca J., Wang R. Y., Alter H. J., Shih J. W. Clinical correlation and genetic polymorphism of the human immunodeficiency virus proviral DNA obtained after polymerase chain reaction amplification. J Infect Dis. 1990 Nov;162(5):1025–1030. doi: 10.1093/infdis/162.5.1025. [DOI] [PubMed] [Google Scholar]
  9. Hardy D. A., Arvin A. M., Yasukawa L. L., Bronzan R. N., Lewinsohn D. M., Hensleigh P. A., Prober C. G. Use of polymerase chain reaction for successful identification of asymptomatic genital infection with herpes simplex virus in pregnant women at delivery. J Infect Dis. 1990 Nov;162(5):1031–1035. doi: 10.1093/infdis/162.5.1031. [DOI] [PubMed] [Google Scholar]
  10. Higuchi R., von Beroldingen C. H., Sensabaugh G. F., Erlich H. A. DNA typing from single hairs. Nature. 1988 Apr 7;332(6164):543–546. doi: 10.1038/332543a0. [DOI] [PubMed] [Google Scholar]
  11. Horsburgh C. R., Jr, Ou C. Y., Jason J., Holmberg S. D., Lifson A. R., Moore J. L., Ward J. W., Seage G. R., Mayer K. H., Evatt B. L. Concordance of polymerase chain reaction with human immunodeficiency virus antibody detection. J Infect Dis. 1990 Aug;162(2):542–545. doi: 10.1093/infdis/162.2.542. [DOI] [PubMed] [Google Scholar]
  12. Jensen J. S., Uldum S. A., Søndergård-Andersen J., Vuust J., Lind K. Polymerase chain reaction for detection of Mycoplasma genitalium in clinical samples. J Clin Microbiol. 1991 Jan;29(1):46–50. doi: 10.1128/jcm.29.1.46-50.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnson P. J., d'Oliveira C. E., Gorrell T. E., Müller M. Molecular analysis of the hydrogenosomal ferredoxin of the anaerobic protist Trichomonas vaginalis. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6097–6101. doi: 10.1073/pnas.87.16.6097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kiviat N. B., Paavonen J. A., Brockway J., Critchlow C. W., Brunham R. C., Stevens C. E., Stamm W. E., Kuo C. C., DeRouen T., Holmes K. K. Cytologic manifestations of cervical and vaginal infections. I. Epithelial and inflammatory cellular changes. JAMA. 1985 Feb 15;253(7):989–996. [PubMed] [Google Scholar]
  15. Krieger J. N., Holmes K. K., Spence M. R., Rein M. F., McCormack W. M., Tam M. R. Geographic variation among isolates of Trichomonas vaginalis: demonstration of antigenic heterogeneity by using monoclonal antibodies and the indirect immunofluorescence technique. J Infect Dis. 1985 Nov;152(5):979–984. doi: 10.1093/infdis/152.5.979. [DOI] [PubMed] [Google Scholar]
  16. Krieger J. N., Tam M. R., Stevens C. E., Nielsen I. O., Hale J., Kiviat N. B., Holmes K. K. Diagnosis of trichomoniasis. Comparison of conventional wet-mount examination with cytologic studies, cultures, and monoclonal antibody staining of direct specimens. JAMA. 1988 Feb 26;259(8):1223–1227. doi: 10.1001/jama.259.8.1223. [DOI] [PubMed] [Google Scholar]
  17. Krieger J. N. Urologic aspects of trichomoniasis. Invest Urol. 1981 May;18(8):411–417. [PubMed] [Google Scholar]
  18. Krieger J. N., Wolner-Hanssen P., Stevens C., Holmes K. K. Characteristics of Trichomonas vaginalis isolates from women with and without colpitis macularis. J Infect Dis. 1990 Feb;161(2):307–311. doi: 10.1093/infdis/161.2.307. [DOI] [PubMed] [Google Scholar]
  19. Leblanc D. J., Lee L. N. Rapid screening procedure for detection of plasmids in streptococci. J Bacteriol. 1979 Dec;140(3):1112–1115. doi: 10.1128/jb.140.3.1112-1115.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Li H. H., Gyllensten U. B., Cui X. F., Saiki R. K., Erlich H. A., Arnheim N. Amplification and analysis of DNA sequences in single human sperm and diploid cells. Nature. 1988 Sep 29;335(6189):414–417. doi: 10.1038/335414a0. [DOI] [PubMed] [Google Scholar]
  21. MANDEL M., HONIGBERG B. M. ISOLATION AND CHARACTERIZATION OF DEOXYRIBONUCLEIC ACID OF TWO SPECIES OF TRICHOMONAS DONNE. J Protozool. 1964 Feb;11:114–116. doi: 10.1111/j.1550-7408.1964.tb01730.x. [DOI] [PubMed] [Google Scholar]
  22. Müller M. Biochemical cytology of trichomonad flagellates. I. Subcellular localization of hydrolases, dehydrogenases, and catalase in Tritrichomonas foetus. J Cell Biol. 1973 May;57(2):453–474. doi: 10.1083/jcb.57.2.453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Müller M. Energy metabolism of protozoa without mitochondria. Annu Rev Microbiol. 1988;42:465–488. doi: 10.1146/annurev.mi.42.100188.002341. [DOI] [PubMed] [Google Scholar]
  24. Ostergaard L., Birkelund S., Christiansen G. Use of polymerase chain reaction for detection of Chlamydia trachomatis. J Clin Microbiol. 1990 Jun;28(6):1254–1260. doi: 10.1128/jcm.28.6.1254-1260.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. PAPANICOLAOU G. N., WOLINSKA W. H. Vaginal cytology in Trichomonas infestation. Int Rec Med Gen Pract Clin. 1955 Sep;168(9):551–556. [PubMed] [Google Scholar]
  26. Riley D. E., Goldman M. A., Gartler S. M. Nucleotide sequence of the 3' nuclease-sensitive region of the human phosphoglycerate kinase 1 (PGK1) gene. Genomics. 1991 Sep;11(1):212–214. doi: 10.1016/0888-7543(91)90121-t. [DOI] [PubMed] [Google Scholar]
  27. Riley D. E., Samadpour M., Krieger J. N. Detection of variable DNA repeats in diverse eukaryotic microorganisms by a single set of polymerase chain reaction primers. J Clin Microbiol. 1991 Dec;29(12):2746–2751. doi: 10.1128/jcm.29.12.2746-2751.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Riley D. E. Very rapid nucleotide sequence analysis of improved, double-stranded minipreps. Gene. 1989 Jan 30;75(1):193–196. doi: 10.1016/0378-1119(89)90396-x. [DOI] [PubMed] [Google Scholar]
  29. Roberts M. C. Characterization of the Tet M determinants in urogenital and respiratory bacteria. Antimicrob Agents Chemother. 1990 Mar;34(3):476–478. doi: 10.1128/aac.34.3.476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Roberts M. C. Characterization of the Tet M determinants in urogenital and respiratory bacteria. Antimicrob Agents Chemother. 1990 Mar;34(3):476–478. doi: 10.1128/aac.34.3.476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rothenberg R. B., Simon R., Chipperfield E., Catterall R. D. Efficacy of selected diagnostic tests for sexually transmitted diseases. JAMA. 1976 Jan 5;235(1):49–51. [PubMed] [Google Scholar]
  32. Rowley A. H., Whitley R. J., Lakeman F. D., Wolinsky S. M. Rapid detection of herpes-simplex-virus DNA in cerebrospinal fluid of patients with herpes simplex encephalitis. Lancet. 1990 Feb 24;335(8687):440–441. doi: 10.1016/0140-6736(90)90667-t. [DOI] [PubMed] [Google Scholar]
  33. Su-Lin K. E., Honigberg B. M. Antigenic analysis of Trichomonas vaginalis strains by quantitative fluorescent antibody methods. Z Parasitenkd. 1983;69(2):161–181. doi: 10.1007/BF00926952. [DOI] [PubMed] [Google Scholar]
  34. Torian B. E., Connelly R. J., Stephens R. S., Stibbs H. H. Specific and common antigens of Trichomonas vaginalis detected by monoclonal antibodies. Infect Immun. 1984 Jan;43(1):270–275. doi: 10.1128/iai.43.1.270-275.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Turner G., Müller M. Failure to detect extranuclear DNA in Trichomonas vaginalis and Tritrichomonas foetus. J Parasitol. 1983 Feb;69(1):234–236. [PubMed] [Google Scholar]
  36. Wachtel E. G., Hudson E. A. The usefulness of cytology. Br J Hosp Med. 1980 Mar;23(3):256-8, 260, 262-5. [PubMed] [Google Scholar]
  37. Wang A. L., Wang C. C. Isolation and characterization of DNA from Tritrichomonas foetus and Trichomonas vaginalis. Mol Biochem Parasitol. 1985 Mar;14(3):323–335. doi: 10.1016/0166-6851(85)90060-x. [DOI] [PubMed] [Google Scholar]
  38. Wølner-Hanssen P., Krieger J. N., Stevens C. E., Kiviat N. B., Koutsky L., Critchlow C., DeRouen T., Hillier S., Holmes K. K. Clinical manifestations of vaginal trichomoniasis. JAMA. 1989 Jan 27;261(4):571–576. doi: 10.1001/jama.1989.03420040109029. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES