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Specific detection and identification of African trypanosomes in bovine peripheral blood by means of a PCR-ELISA assay

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dc.contributor.author Cabrera, L.
dc.contributor.author De Witte, J.
dc.contributor.author Victor, B.
dc.contributor.author Vermeiren, L.
dc.contributor.author Zimic, M.
dc.contributor.author Brandt, J.
dc.contributor.author Geysen, D.
dc.date.accessioned 2009-12-18T12:50:50Z
dc.date.available 2009-12-18T12:50:50Z
dc.date.issued 2009
dc.identifier.issn 0304-4017
dc.identifier.doi http://dx.doi.org/10.1016/j.vetpar.2009.06.017
dc.identifier.other ITG-A1B
dc.identifier.other ITG-A2B
dc.identifier.other ITG-A3B
dc.identifier.other ITG-X6A
dc.identifier.other ITG-ALA
dc.identifier.other ANIMAL
dc.identifier.other U-ANIMAL
dc.identifier.other JIF
dc.identifier.other DOI
dc.identifier.other UPD17
dc.identifier.other ABSTRACT
dc.identifier.uri http://hdl.handle.net/10390/2847
dc.description.abstract The aim of the present study was to develop a PCR-ELISA assay for the detection and differentiation of the main African pathogen trypanosomal species present in peripheral blood of cattle. The proposed methodology allows to specifically differentiate Trypanosoma congolense, Trypanosoma vivax and the subgenus Trypanozoon, by means of a sensitive universal PCR amplifying trypanosome DNA followed by an ELISA-based hybridization with three highly specific probes. The semi-nested PCR had a sensitivity of 15 fg, 15 fg, and 0.15 fg of DNA from T. vivax, T. congolense, and Trypanosoma brucei brucei, respectively that is sufficient to detect parasites in blood during the chronic phase of the disease. Biotinylated second round asymmetric PCR amplification products were used in an ELISA set up using three species-specific probes for the diagnosis of T. congolense (type Riverine, Kilifi or Savannah), T. vivax and T. brucei brucei. A factor O.D. of 0.082 was determined on blood samples from bovines (n=18) from a non-endemic area in Africa. In a pilot study of blood samples of naturally and experimentally Trypanosoma infected cattle previously characterized by PCR-RFLP (n=42), a high rate of concordance (93.3%) was found between PCR-RFLP and PCR-ELISA. There is a good ratio between positive and negative O.D. values (3.00 vs. 0.1) and the technique can also be used to distinguish mixed infections. en
dc.language English en
dc.subject Animal diseases en
dc.subject Bovines en
dc.subject Protozoal diseases en
dc.subject Trypanosomiasis, African en
dc.subject Trypanosoma congolense en
dc.subject Trypanosoma brucei en
dc.subject Trypanosoma vivax en
dc.subject Trypanozoon en
dc.subject Differential diagnosis en
dc.subject Assays en
dc.subject PCR-ELISA en
dc.title Specific detection and identification of African trypanosomes in bovine peripheral blood by means of a PCR-ELISA assay en
dc.type Article en
dc.citation.issue 2-4 en
dc.citation.jtitle Veterinary Parasitology en
dc.citation.volume 164 en
dc.citation.pages 111-117 en
dc.identifier.pmid http://www.ncbi.nlm.nih.gov/pubmed/19619947
dc.citation.jabbreviation Vet Parasitol en


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