Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 May;33(5):1308–1313. doi: 10.1128/jcm.33.5.1308-1313.1995

PCR-based study of conserved and variable DNA sequences of Tritrichomonas foetus isolates from Saskatchewan, Canada.

D E Riley 1, B Wagner 1, L Polley 1, J N Krieger 1
PMCID: PMC228151  PMID: 7615746

Abstract

The protozoan parasite Tritrichomonas foetus causes infertility and spontaneous abortion in cattle. In Saskatchewan, Canada, the culture prevalence of trichomonads was 65 of 1,048 (6%) among 1,048 bulls tested within a 1-year period ending in April 1994. Saskatchewan was previously thought to be free of the parasite. To confirm the culture results, possible T. foetus DNA presence was determined by the PCR. All of the 16 culture-positive isolates tested were PCR positive by a single-band test, but one PCR product was weak. DNA fingerprinting by both T17 PCR and randomly amplified polymorphic DNA PCR revealed genetic variation or polymorphism among the T. foetus isolates. T17 PCR also revealed conserved loci that distinguished these T. foetus isolates from Trichomonas vaginalis, from a variety of other protozoa, and from prokaryotes. TCO-1 PCR, a PCR test designed to sample DNA sequence homologous to the 5' flank of a highly conserved cell division control gene, detected genetic polymorphism at low stringency and a conserved, single locus at higher stringency. These findings suggested that T. foetus isolates exhibit both conserved genetic loci and polymorphic loci detectable by independent PCR methods. Both conserved and polymorphic genetic loci may prove useful for improved clinical diagnosis of T. foetus. The polymorphic loci detected by PCR suggested either a long history of infection or multiple lines of T. foetus infection in Saskatchewan. Polymorphic loci detected by PCR may provide data for epidemiologic studies of T. foetus.

Full Text

The Full Text of this article is available as a PDF (275.5 KB).

Selected References

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

  1. Bostock A., Khattak M. N., Matthews R., Burnie J. Comparison of PCR fingerprinting, by random amplification of polymorphic DNA, with other molecular typing methods for Candida albicans. J Gen Microbiol. 1993 Sep;139(9):2179–2184. doi: 10.1099/00221287-139-9-2179. [DOI] [PubMed] [Google Scholar]
  2. Burgess D. E. Clonal and geographic distribution of a surface antigen of Tritrichomonas foetus. J Protozool. 1988 Feb;35(1):119–122. doi: 10.1111/j.1550-7408.1988.tb04090.x. [DOI] [PubMed] [Google Scholar]
  3. Campbell J., Schumann F., Janzen E., Wagner B., Polley L. Saskatchewan. Trichomonas fetus in bulls in Saskatchewan. Can Vet J. 1993 Sep;34(9):571–572. [PMC free article] [PubMed] [Google Scholar]
  4. Carnaby S., Katelaris P. H., Naeem A., Farthing M. J. Genotypic heterogeneity within Giardia lamblia isolates demonstrated by M13 DNA fingerprinting. Infect Immun. 1994 May;62(5):1875–1880. doi: 10.1128/iai.62.5.1875-1880.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Flint J., Boyce A. J., Martinson J. J., Clegg J. B. Population bottlenecks in Polynesia revealed by minisatellites. Hum Genet. 1989 Oct;83(3):257–263. doi: 10.1007/BF00285167. [DOI] [PubMed] [Google Scholar]
  6. Gill P., Jeffreys A. J., Werrett D. J. Forensic application of DNA 'fingerprints'. Nature. 1985 Dec 12;318(6046):577–579. doi: 10.1038/318577a0. [DOI] [PubMed] [Google Scholar]
  7. Ho M. S., Conrad P. A., Conrad P. J., LeFebvre R. B., Perez E., BonDurant R. H. Detection of bovine trichomoniasis with a specific DNA probe and PCR amplification system. J Clin Microbiol. 1994 Jan;32(1):98–104. doi: 10.1128/jcm.32.1.98-104.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hodgson J. L., Jones D. W., Widders P. R., Corbeil L. B. Characterization of Tritrichomonas foetus antigens by use of monoclonal antibodies. Infect Immun. 1990 Sep;58(9):3078–3083. doi: 10.1128/iai.58.9.3078-3083.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jeffreys A. J., Neumann R., Wilson V. Repeat unit sequence variation in minisatellites: a novel source of DNA polymorphism for studying variation and mutation by single molecule analysis. Cell. 1990 Feb 9;60(3):473–485. doi: 10.1016/0092-8674(90)90598-9. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Kimura H., Shibata M., Kuzushima K., Nishikawa K., Nishiyama Y., Morishima T. Detection and direct typing of herpes simplex virus by polymerase chain reaction. Med Microbiol Immunol. 1990;179(4):177–184. doi: 10.1007/BF00195248. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. McCool C. J., Townsend M. P., Wolfe S. G., Simpson M. A., Olm T. C., Jayawardhana G. A., Carney J. V. Prevalence of bovine venereal disease in the Victoria River District of the Northern Territory: likely economic effects and practicable control measures. Aust Vet J. 1988 May;65(5):153–156. doi: 10.1111/j.1751-0813.1988.tb14445.x. [DOI] [PubMed] [Google Scholar]
  14. Mickelsen W. D., Paisley L. G., Anderson P. B. Prevalence of postservice pyometra in a herd of beef cows infected with trichomoniasis: A case report. Theriogenology. 1986 May;25(5):741–744. doi: 10.1016/0093-691x(86)90132-9. [DOI] [PubMed] [Google Scholar]
  15. Quon D. V., Delgadillo M. G., Khachi A., Smale S. T., Johnson P. J. Similarity between a ubiquitous promoter element in an ancient eukaryote and mammalian initiator elements. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4579–4583. doi: 10.1073/pnas.91.10.4579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rae D. O. Impact of trichomoniasis on the cow-calf producer's profitability. J Am Vet Med Assoc. 1989 Mar 15;194(6):771–775. [PubMed] [Google Scholar]
  17. Rhyan J. C., Stackhouse L. L., Quinn W. J. Fetal and placental lesions in bovine abortion due to Tritrichomonas foetus. Vet Pathol. 1988 Sep;25(5):350–355. doi: 10.1177/030098588802500503. [DOI] [PubMed] [Google Scholar]
  18. Riley D. E., Campbell L. A., Puolakkainen M., Krieger J. N. Trichomonas vaginalis and early evolving DNA and protein sequences of the CDC2/28 protein kinase family. Mol Microbiol. 1993 May;8(3):517–519. doi: 10.1111/j.1365-2958.1993.tb01595.x. [DOI] [PubMed] [Google Scholar]
  19. Riley D. E., Krieger J. N. Rapid and practical DNA isolation from Trichomonas vaginalis and other nuclease-rich protozoa. Mol Biochem Parasitol. 1992 Mar;51(1):161–163. doi: 10.1016/0166-6851(92)90212-3. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Ross B. C., Raios K., Jackson K., Sievers A., Dwyer B. Differentiation of Mycobacterium tuberculosis strains by use of a nonradioactive Southern blot hybridization method. J Infect Dis. 1991 Apr;163(4):904–907. doi: 10.1093/infdis/163.4.904. [DOI] [PubMed] [Google Scholar]
  22. Stevens D. A., Odds F. C., Scherer S. Application of DNA typing methods to Candida albicans epidemiology and correlations with phenotype. Rev Infect Dis. 1990 Mar-Apr;12(2):258–266. doi: 10.1093/clinids/12.2.258. [DOI] [PubMed] [Google Scholar]
  23. Torian B. E., Connelly R. J., Barnes R. C., Kenny G. E. Antigenic heterogeneity in the 115,000 Mr major surface antigen of Trichomonas vaginalis. J Protozool. 1988 May;35(2):273–280. doi: 10.1111/j.1550-7408.1988.tb04343.x. [DOI] [PubMed] [Google Scholar]
  24. Wahls W. P., Wallace L. J., Moore P. D. Hypervariable minisatellite DNA is a hotspot for homologous recombination in human cells. Cell. 1990 Jan 12;60(1):95–103. doi: 10.1016/0092-8674(90)90719-u. [DOI] [PubMed] [Google Scholar]
  25. Wang G., Whittam T. S., Berg C. M., Berg D. E. RAPD (arbitrary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains. Nucleic Acids Res. 1993 Dec 25;21(25):5930–5933. doi: 10.1093/nar/21.25.5930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wong Z., Wilson V., Jeffreys A. J., Thein S. L. Cloning a selected fragment from a human DNA 'fingerprint': isolation of an extremely polymorphic minisatellite. Nucleic Acids Res. 1986 Jun 11;14(11):4605–4616. doi: 10.1093/nar/14.11.4605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wöstemeyer J. Strain-dependent variation in ribosomal DNA arrangement in Absidia glauca. Eur J Biochem. 1985 Jan 15;146(2):443–448. doi: 10.1111/j.1432-1033.1985.tb08671.x. [DOI] [PubMed] [Google Scholar]
  28. Yule A., Skirrow S. Z., Bonduran R. H. Bovine trichomoniasis. Parasitol Today. 1989 Dec;5(12):373–377. doi: 10.1016/0169-4758(89)90298-6. [DOI] [PubMed] [Google Scholar]
  29. van Belkum A., Homan W., Limper L., Quint W. G. Genotyping isolates and clones of Giardia duodenalis by polymerase chain reaction: implications for the detection of genetic variation among protozoan parasite species. Mol Biochem Parasitol. 1993 Sep;61(1):69–77. doi: 10.1016/0166-6851(93)90159-u. [DOI] [PubMed] [Google Scholar]

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

RESOURCES