Journal article
Fluorescence in situ hybridization for identification of Tritrichomonas foetus in formalin-fixed and paraffin-embedded histological specimens of intestinal trichomonosis
Veterinary parasitology, Vol.172(1), pp.139-143
2010
DOI: 10.1016/j.vetpar.2010.04.014
PMID: 20447769
Abstract
In the present study a highly species-specific oligonucleotide sequence of
Tritrichomonas foetus 18S rRNA was used to design an antisense probe for identification of
T. foetus in formalin-fixed and paraffin-embedded histological specimens by means of fluorescence in situ hybridization (FISH). Using archival histological specimens from several species with light microscopic evidence of intestinal trichomonosis, and under optimized hybridization conditions, the probe positively identified trichomonads in colonic specimens from piglets and a kitten with PCR-confirmed
T. foetus infection. Neither positive hybridization of the probe or PCR amplification of
T. foetus DNA was observed in histological specimens from hamster (
Tritrichomonas muris), turkey, nor mouse (
Entamoeba muris) intestinal protozoal infections. Sequence-specific binding of the probe was further verified by successfully out-competing the hybridization with 10
×
molar excess unlabeled probe and failure of a labeled sense probe to hybridize. The FISH assay described here enables simultaneous location and molecular identification of
T. foetus in formalin-fixed and paraffin-embedded histological specimens of intestinal trichomonosis. The methods employed are likely to also be applicable to probes designed for specific recognition of other trichomonad species, especially in mammalian tissue where red blood cell auto-fluorescence can be easily differentiated from the hybridization signal of trichomonads.
Details
- Title: Subtitle
- Fluorescence in situ hybridization for identification of Tritrichomonas foetus in formalin-fixed and paraffin-embedded histological specimens of intestinal trichomonosis
- Creators
- J.L Gookin - Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, United StatesM.R Stone - Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, United StatesM.J Yaeger - Veterinary Pathology, College of Veterinary Medicine, Iowa State University, Ames, IA, United StatesD.K Meyerholz - Department of Pathology, University of Iowa Carver College of Medicine, Iowa City, IA, United StatesPeter Moisan - Rollins Animal Disease Diagnostic Laboratory, Raleigh, NC, United States
- Resource Type
- Journal article
- Publication Details
- Veterinary parasitology, Vol.172(1), pp.139-143
- DOI
- 10.1016/j.vetpar.2010.04.014
- PMID
- 20447769
- NLM abbreviation
- Vet Parasitol
- ISSN
- 0304-4017
- eISSN
- 1873-2550
- Publisher
- Elsevier B.V
- Language
- English
- Date published
- 2010
- Academic Unit
- Pathology
- Record Identifier
- 9984083858702771
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