title: Design and validation of an oligonucleotide probe for the detection of protozoa from the order Trichomonadida using chromogenic in situ hybridization.
abstract: Infections with protozoal parasites of the order Trichomonadida are often observed in veterinary medicine. Based on the trichomonad species involved these infections are either asymptomatic or can lead to sometimes serious disease. To further study protozoal agents of the order Trichomonadida the establishment of a method to detect trichomonads directly in the tissue, allowing parasite-lesion correlation, is necessary. Here we describe the design and evaluation of an oligonucleotide probe for chromogenic in situ hybridization, theoretically allowing detection of all hitherto known members of the order Trichomonadida. The probe was designed on a region of the 18S ribosomal RNA gene homologue for all representatives of the order Trichomonadida available in the GenBank. Functionality of the probe was proven using protozoal cultures containing different trichomonads (Monocercomonas colubrorum, Hypotrichomonas acosta, Pentatrichomonas hominis, Trichomitus batrachorum, Trichomonas gallinae, Tetratrichomonas gallinarum, Tritrichomonas foetus, and Tritrichomonas augusta). Furthermore, three different tissue sections containing either T. gallinae, T. foetus or Histomonas meleagridis were tested positive. Additionally, to rule out cross-reactivity of the probe a large number of different pathogenic protozoal agents, fungi, bacteria and viruses were tested and gave negative results. The probe presented here can be considered an important tool for diagnosis of all to date described relevant protozoal parasites of the order Trichomonadida in tissue samples.
authors.affiliation: Institute of Pathology and Forensic Veterinary Medicine, Department of Pathobiology, University of Veterinary Medicine, Veterinärplatz 1, 1210 Vienna, Austria.
authors.forename: Meike Marissa
authors.initials: MM
authors.lastname: Mostegl
authors.forename: Barbara
authors.initials: B
authors.lastname: Richter
authors.forename: Nora
authors.initials: N
authors.lastname: Nedorost
authors.forename: Anton
authors.initials: A
authors.lastname: Maderner
authors.forename: Nora
authors.initials: N
authors.lastname: Dinhopl
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Kulda
authors.forename: Dieter
authors.initials: D
authors.lastname: Liebhart
authors.forename: Michael
authors.initials: M
authors.lastname: Hess
authors.forename: Herbert
authors.initials: H
authors.lastname: Weissenböck
mesh_terms: D000818:Animals; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D005767:Gastrointestinal Diseases / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D017403:In Situ Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction; D012337:RNA, Ribosomal, 18S / Q000737:chemistry / Q000235:genetics; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000469:parasitology*; D016834:Trichomonadida / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D016054:DNA, Protozoan; D015345:Oligonucleotide Probes; D012337:RNA, Ribosomal, 18S
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