Abstract
The application of nucleic acid hybridization techniques in the identification of most protozoan parasites, using species-specific DNA probes, has recently been described by several investigators. Species-specific DNA probes have been employed in the characterization of trypanosome infections in cattle and tsetse from Uganda. Most infections revealed by our DNA probes were mixed. Using these probes, a mixed infection with Trypanosoma brucei, T. vivax and both savannah and Kilifi types of T. congolense was revealed in one cow. This mixed infection could not have been detected by any of the classical parasitological methods. Isolates made from natural field infections, which had been passaged in laboratory animals, were found to consist of homogeneous trypanosome species. This was demonstrated in all of 47 stabilates which were homogeneous infections either of savannah type T. congolense or T. brucei.
The method of sample preparation for DNA probe analysis was modified to suit field conditions. The samples, which were spot-blotted onto nylon niters and either immediately denatured or left undenatured, could be kept at room temperature for 1 month with only a moderate loss of hybridization signal intensities. Although hybridization signals were visible in undenatured samples, those seen with the samples that had been denatured were clearly more intense. This approach eliminates the need for liquid nitrogen and/or an incubator in the field. The simplicity, sensitivity and specificity of this diagnostic technique using species-specific DNA probes, make it an important tool for future studies of the epidemiology of African trypanosomiases.
Résumé
L’emploi de techniques d’hybridation des acides nucléiques pour identifier la plupart des parasites protozoaires a été récemment décrit par plusieurs groupes. Des sondes ADN spécifiques de l’espèce ont été utilisées pour caractériser des infections dues aux trypanosomes chez les bovins et les mouches tsé-tsé en provenance de l’Ouganda. La plupart des infections étaient mixtes: par exemple, chez une vâche, une infection mixte par Trypanosome brucei, T. vivax ainsi que par T. congolense du type Kilifi et des savannes. Cette infection mixte n’aurait pas pu être détectée par les méthodes classiques de parasitologic. 47 isolats obtenus d’infections dans la nature et passés dans des animaux de laboratoire ont tous contenu des espèce homogénes, de type savanne T. brucei ou T. congolense.
La préparation d’échantillons était modifiée pour tenir compte des conditions sur le terrain. Les échantillons, spottés sur des filtres nylon en double, dont un soumis à dénaturation, pouvaient être maintenus à temperature ambiente pendant 1 mois avec seulement une perte modérée d’intensité du signal. Les signaux obtenus avec les échantillons dénaturés étaient plus intense que ceux obtenus avec les échantillons non-dénaturés. Cette approche supprime le besoin de recourir à l’azote liquide et/ou d’un incubateur sur le terrain. La simplicité, la sensibilité et la spécificité de cette technique diagnostique basée sur l’utilisation de sondes ADN spécifiques de l’espèce en font un outil important pour les futures études d’épidemiologie de la trypanosomiase africaine.
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Nyeko, J.H.P., Ole-Moiyoi, O.K., Majiwa, P.A.O. et al. Characterization of Trypanosome Isolates from Cattle in Uganda using Species-Specific DNA Probes Reveals Predominance of Mixed Infections. Int J Trop Insect Sci 11, 271–280 (1990). https://doi.org/10.1017/S1742758400012674
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DOI: https://doi.org/10.1017/S1742758400012674