PMID- 16920265
OWN - NLM
STAT- MEDLINE
DCOM- 20080731
LR  - 20091119
IS  - 0304-4017 (Print)
IS  - 0304-4017 (Linking)
VI  - 142
IP  - 3-4
DP  - 2006 Dec 20
TI  - PCR for the identification and differentiation of Histomonas meleagridis, 
      Tetratrichomonas gallinarum and Blastocystis spp.
PG  - 223-30
AB  - In the present investigation PCR assays were developed for the rapid detection 
      and differentiation of two poultry flagellates: Histomonas meleagridis and 
      Tetratrichomonas gallinarum as well as the protozoan microorganism: Blastocystis 
      spp. The nucleotide sequences of the small subunit ribosomal RNAs were used for 
      primer construction obtaining fragments which vary in size for each 
      microorganism. The established PCRs were able to detect DNA obtained from one 
      microorganism of T. gallinarum and Blastocystis spp. propagated in vitro, proving 
      the high analytical sensitivity of the method. DNA isolated from 10 protozoa was 
      sufficient to detect H. meleagridis. To assess specificity, each PCR assay was 
      performed with DNA from either H. meleagridis and/or T. gallinarum and/or 
      Blastocystis spp. as well as with DNA from several other protozoan parasites 
      (Eimeria tenella, Toxoplasma gondii, Cryptosporidia spp., Trichomonas gallinae, 
      Entamoeba invadens, Entamoeba ranarum), fungi (Aspergillus fumigatus, Candida 
      albicans), bacteria (Staphylococcae, Streptococcae, E. coli, Clostridium 
      perfringens, Camplyobacter jejuni, Proteus) and viruses (fowl adenovirus serotype 
      4, avian reovirus) as well as livers and caecal samples from turkeys and 
      specified pathogen free (spf) chickens. No cross-reactions with any of these 
      samples were observed with the primer sets for the detection of H. meleagridis 
      and Blastocystis spp. The primers designed for the identification of T. 
      gallinarum yielded a PCR product with DNA of Trichomonas gallinae that had the 
      identical size as the amplicon obtained with DNA from T. gallinarum. However, no 
      PCR products resulted from any of the other samples tested with these primers. 
      Liver and caecal samples from turkeys and chickens from flocks with outbreaks of 
      histomonosis also named as "histomoniasis" originating from geographically 
      distinct regions were investigated with the established PCRs. This is also the 
      first report about the detection of the nucleic acid of H. meleagridis, T. 
      gallinarum and Blastocystis spp. nucleic acid in the livers and/or caeca of 
      laying hens and turkeys obtained from field outbreaks. Hence, the established PCR 
      assays proved to be a rapid and sensitive diagnostic tool for the direct 
      detection and differentiation of H. meleagridis, T. gallinarum and Blastocystis 
      spp. nucleic acid in organ samples of infected turkeys and chickens regardless of 
      the geographic origin.
FAU - Grabensteiner, E
AU  - Grabensteiner E
AD  - Department for Farm Animals and Herd Management, Clinic for Avian, Reptile and 
      Fish Medicine, University of Veterinary Medicine Vienna, Veterinarplatz 1, 1210 
      Vienna, Austria.
FAU - Hess, M
AU  - Hess M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060821
PL  - Netherlands
TA  - Vet Parasitol
JT  - Veterinary parasitology
JID - 7602745
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Protozoan)
RN  - 0 (DNA, Ribosomal)
SB  - IM
MH  - Animals
MH  - Blastocystis/classification/genetics/isolation & purification
MH  - Chickens/parasitology
MH  - DNA Primers/chemistry
MH  - DNA, Protozoan/chemistry/isolation & purification
MH  - DNA, Ribosomal/*chemistry/isolation & purification
MH  - Eukaryota/classification/genetics/*isolation & purification
MH  - Liver/parasitology
MH  - Polymerase Chain Reaction/methods/*veterinary
MH  - Poultry Diseases/*diagnosis/parasitology
MH  - Protozoan Infections/diagnosis
MH  - *Protozoan Infections, Animal
MH  - Sensitivity and Specificity
MH  - Species Specificity
MH  - Specific Pathogen-Free Organisms
MH  - Trichomonadida/classification/genetics/isolation & purification
MH  - Turkeys/parasitology
EDAT- 2006/08/22 09:00
MHDA- 2008/08/01 09:00
CRDT- 2006/08/22 09:00
PHST- 2005/01/20 00:00 [received]
PHST- 2006/07/03 00:00 [revised]
PHST- 2006/07/13 00:00 [accepted]
PHST- 2006/08/22 09:00 [pubmed]
PHST- 2008/08/01 09:00 [medline]
PHST- 2006/08/22 09:00 [entrez]
AID - S0304-4017(06)00409-2 [pii]
AID - 10.1016/j.vetpar.2006.07.011 [doi]
PST - ppublish
SO  - Vet Parasitol. 2006 Dec 20;142(3-4):223-30. doi: 10.1016/j.vetpar.2006.07.011. 
      Epub 2006 Aug 21.