Design and validation of an oligonucleotide probe for the detection of protozoa from the order Trichomonadida using chromogenic in situ hybridization.
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.
Mostegl
Meike Marissa
MM
Institute of Pathology and Forensic Veterinary Medicine, Department of Pathobiology, University of Veterinary Medicine, Veterinärplatz 1, 1210 Vienna, Austria.
Richter
Barbara
B
Nedorost
Nora
N
Maderner
Anton
A
Dinhopl
Nora
N
Kulda
Jaroslav
J
Liebhart
Dieter
D
Hess
Michael
M
Weissenböck
Herbert
H
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*
D016054:DNA, Protozoan; D015345:Oligonucleotide Probes; D012337:RNA, Ribosomal, 18S
Virology. 2004 Oct 25;328(2):301-10
15464850
Parasitol Res. 2009 Sep;105(3):751-6
19421777
Protist. 2007 Jul;158(3):365-83
17499022
J Comp Pathol. 2008 Feb-Apr;138(2-3):63-71
18295780
Vet Parasitol. 2006 Dec 20;142(3-4):223-30
16920265
J Parasitol. 2003 Feb;89(1):99-104
12659310
J Comp Pathol. 2006 Feb-Apr;134(2-3):211-8
16615936
J Eukaryot Microbiol. 2002 Mar-Apr;49(2):154-63
12046599
J Comp Pathol. 1976 Jan;86(1):59-66
1254746
J Comp Pathol. 2006 Nov;135(4):237-42
17069832
Vet Parasitol. 2007 Apr 30;145(3-4):349-51
17314011
Am J Vet Res. 2007 Jul;68(7):783-7
17605615
Mol Phylogenet Evol. 2009 Jul;52(1):217-24
19306937
Parasitol Res. 2008 Nov;103(6):1311-6
18682985
Int J Syst Evol Microbiol. 2006 Jan;56(Pt 1):305-12
16403903
J Am Vet Med Assoc. 2003 May 15;222(10):1376-9
12762381
Exp Parasitol. 1954 Jul;3(4):368-402
13183096
Int J Parasitol. 2004 Jul;34(8):963-70
15217735