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Ribosomal DNA analysis of tsetse and non-tsetse transmitted Ethiopian Trypanosoma vivax strains in view of improved molecular diagnosis

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Show simple item record Fikru, R. Matetovici, I. Rogé, S. Merga, B. Goddeeris, B. M. Büscher, P. van Reet, N. 2016-06-03T12:42:02Z 2016-06-03T12:42:02Z 2016
dc.identifier.issn 0304-4017
dc.identifier.other ITG-B1B
dc.identifier.other ITG-B2B
dc.identifier.other ITG-B3B
dc.identifier.other ITG-B6A
dc.identifier.other ITG-BLB
dc.identifier.other DBM
dc.identifier.other U-PARDIA
dc.identifier.other JIF
dc.identifier.other DOI
dc.identifier.other FTA
dc.identifier.other OAA
dc.identifier.other Abstract
dc.identifier.other UPD62
dc.description.abstract Animal trypanosomosis caused by Trypanosoma vivax (T. vivax) is a devastating disease causing serious economic losses. Most molecular diagnostics for T. vivax infection target the ribosomal DNA locus (rDNA) but are challenged by the heterogeneity among T. vivax strains. In this study, we investigated the rDNA heterogeneity of Ethiopian T. vivax strains in relation to their presence in tsetse-infested and tsetse-free areas and its effect on molecular diagnosis. We sequenced the rDNA loci of six Ethiopian (three from tsetse-infested and three from tsetse-free areas) and one Nigerian T. vivax strain. We analysed the obtained sequences in silico for primer-mismatches of some commonly used diagnostic PCR assays and for GC content. With these data, we selected some rDNA diagnostic PCR assays for evaluation of their diagnostic accuracy. Furthermore we constructed two phylogenetic networks based on sequences within the smaller subunit (SSU) of 18S and within the 5.8S and internal transcribed spacer 2 (ITS2) to assess the relatedness of Ethiopian T. vivax strains to strains from other African countries and from South America. In silico analysis of the rDNA sequence showed important mismatches of some published diagnostic PCR primers and high GC content of T. vivax rDNA. The evaluation of selected diagnostic PCR assays with specimens from cattle under natural T. vivax challenge showed that this high GC content interferes with the diagnostic accuracy of PCR, especially in cases of mixed infections with T. congolense. Adding betain to the PCR reaction mixture can enhance the amplification of T. vivax rDNA but decreases the sensitivity for T. congolense and Trypanozoon. The networks illustrated that Ethiopian T. vivax strains are considerably heterogeneous and two strains (one from tsetse-infested and one from tsetse-free area) are more related to the West African and South American strains than to the East African strains. The rDNA locus sequence of six Ethiopian T. vivax strains showed important differences and higher GC content compared to other animal trypanosomes but could not be related to their origin from tsetse-infested or tsetse-free area. The high GC content of T. vivax DNA renders accurate diagnosis of all pathogenic animal trypanosomes with one single PCR problematic. en_US
dc.language English en_US
dc.subject Protozoal diseases en_US
dc.subject Animal diseases en_US
dc.subject Nagana en_US
dc.subject Trypanosoma vivax en_US
dc.subject Cattle en_US
dc.subject Vectors en_US
dc.subject Tsetse flies en_US
dc.subject Glossina morsitans morsitans en_US
dc.subject Molecular diagnostic techniques en_US
dc.subject Strains en_US
dc.subject Ribosomal DNA en_US
dc.subject Heterogeneity en_US
dc.subject Phylogenetics en_US
dc.subject Accuracy en_US
dc.subject Amplification en_US
dc.subject Sensitivity en_US
dc.subject Ethiopia en_US
dc.subject Africa, East en_US
dc.title Ribosomal DNA analysis of tsetse and non-tsetse transmitted Ethiopian Trypanosoma vivax strains in view of improved molecular diagnosis en_US
dc.type Article en_US
dc.citation.jtitle Veterinary Parasitology en_US
dc.citation.volume 220 en_US
dc.citation.pages 15-22 en_US
dc.citation.jabbreviation Vet Parasitol en_US

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