An improved molecular assay for Tritrichomonas foetus

Robert A Grahn, Robert Bondurant, K. A. Van Hoosear, R. L. Walker, Leslie A Lyons

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

Tritrichomonas foetus (T. foetus) is the causative agent of bovine trichomonosis, a sexually transmitted disease leading to abortion (from 1 to 8 months gestation), infertility, and occasional pyometra. The annual losses to the U.S. beef industry are estimated to be in the hundreds of millions of dollars. Currently, the "gold standard" diagnostic test for trichomonosis in most countries is the cultivation of live organisms from reproductive secretions. The cultured organisms can then be followed by PCR assays with primers that amplify T. foetus to the exclusion of all other trichomonad species. Thus, negative results present as null data, indistinguishable from failed PCR amplification during T. foetus specific amplification. Our newly developed assay improves previously developed PCR based techniques by using diagnostic size variants from within the internal transcribed spacer 1 (ITS1) region that is between the 18S rRNA and 5.8S rRNA subunits. This new PCR assay amplifies trichomonad DNA from a variety of genera and positively identifies the causative agent in the bovine trichomonad infection. This approach eliminates false negatives found in some current assays as well as identifying the causative agent of trichomonad infection. Additionally, our assay incorporates a fluorescently labeled primer enabling high sensitivity and rapid assessment of the specific trichomonad species. Moreover, electrophoretic separation of amplified samples can be outsourced, thus eliminating the need for diagnostic laboratories to purchase expensive analysis equipment.

Original languageEnglish (US)
Pages (from-to)33-41
Number of pages9
JournalVeterinary Parasitology
Volume127
Issue number1
DOIs
StatePublished - Jan 4 2005

Keywords

  • Polymerase chain reaction (PCR)
  • Trichomoniasis
  • Trichomonosis
  • Tritrichomonas foetus (T. foetus)

ASJC Scopus subject areas

  • Parasitology
  • Medicine(all)
  • veterinary(all)

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