Fluorescence in situ hybridization for identification of Tritrichomonas foetus in formalin-fixed and paraffin-embedded histological specimens of intestinal trichomoniasis
| Autor: | Maria R. Stone, David K. Meyerholz, Michael J. Yaeger, Peter Moisan, Jody L. Gookin |
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| Rok vydání: | 2009 |
| Předmět: |
Swine
Hamster Trichomonas Infections Tritrichomonas foetus Polymerase Chain Reaction law.invention law medicine RNA Ribosomal 18S Animals Polymerase chain reaction In Situ Hybridization Fluorescence General Veterinary biology medicine.diagnostic_test Oligonucleotide Histocytochemistry Histology General Medicine Ribosomal RNA DNA Protozoan biology.organism_classification Molecular biology Intestinal Diseases Parasitology Microscopy Fluorescence Cats Fluorescence in situ hybridization |
| Zdroj: | Veterinary parasitology. 172(1-2) |
| ISSN: | 1873-2550 |
| Popis: | 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 trichomoniasis, 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 x 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 trichomoniasis. 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. |
| Databáze: | OpenAIRE |
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