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

Leyda Cabrera, Jacob De Witte, Björn Victor, Lieve Vermeiren, Mirko Zimic, Jef Brandt, Dirk Geysen

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)111-117
Number of pages7
JournalVeterinary Parasitology
Volume164
Issue number2-4
DOIs
StatePublished - Oct 14 2009
Externally publishedYes

Fingerprint

Trypanosomiasis
Trypanosoma vivax
Trypanosoma congolense
Enzyme-Linked Immunosorbent Assay
Trypanosoma brucei brucei
enzyme-linked immunosorbent assay
Polymerase Chain Reaction
cattle
blood
assays
restriction fragment length polymorphism
Trypanosoma
Restriction Fragment Length Polymorphisms
DNA
mixed infection
savannas
hybridization
parasites
sampling
pathogens

Keywords

  • African bovine trypanosomes
  • Diagnosis
  • PCR-ELISA
  • T. brucei
  • T. congolense
  • T. vivax
  • Trypanozoon

ASJC Scopus subject areas

  • Parasitology
  • veterinary(all)

Cite this

Specific detection and identification of African trypanosomes in bovine peripheral blood by means of a PCR-ELISA assay. / Cabrera, Leyda; De Witte, Jacob; Victor, Björn; Vermeiren, Lieve; Zimic, Mirko; Brandt, Jef; Geysen, Dirk.

In: Veterinary Parasitology, Vol. 164, No. 2-4, 14.10.2009, p. 111-117.

Research output: Contribution to journalArticle

Cabrera, Leyda ; De Witte, Jacob ; Victor, Björn ; Vermeiren, Lieve ; Zimic, Mirko ; Brandt, Jef ; Geysen, Dirk. / Specific detection and identification of African trypanosomes in bovine peripheral blood by means of a PCR-ELISA assay. In: Veterinary Parasitology. 2009 ; Vol. 164, No. 2-4. pp. 111-117.
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