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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Development and evaluation of an ITS1 "Touchdown" PCR for assessment of drug efficacy against animal African trypanosomosis
Veterinary Parasitology, Volume 202, No. 3-4, Year 2014
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Description
Animal African trypanosomoses (AAT) are caused by flagellated protozoa of the Trypanosoma genus and contribute to considerable losses in animal production in Africa, Latin America and South East Asia. Trypanosoma congolense is considered the economically most important species. Drug resistant T. congolense strains present a threat to the control of AAT and have triggered research into discovery of novel trypanocides. In vivo assessment of trypanocidal efficacy relies on monitoring of treated animals with microscopic parasite detection methods. Since these methods have poor sensitivity, follow-up for up to 100 days after treatment is recommended to increase the chance of detecting recurrent parasitaemia waves. Molecular techniques are more amendable to high throughput processing and are generally more sensitive than microscopic detection, thus bearing the potential of shortening the 100-day follow up period. The study presents a "Touchdown" PCR targeting the internal transcribed spacer 1 of the ribosomal DNA (ITS1 TD PCR) that enables detection and discrimination of different Trypanosoma taxa in a single run due to variations in PCR product sizes. The assay achieves analytical sensitivity of 10 parasites per ml of blood for detection of T. congolense savannah type and T. brucei, and 100 parasites per ml of blood for detection of T. vivax in infected mouse blood. The ITS1 TD PCR was evaluated on cattle experimentally infected with T. congolense during an investigational new veterinary trypanocide drug efficacy study. ITS1 TD PCR demonstrated comparable performance to microscopy in verifying trypanocide treatment success, in which parasite DNA became undetectable in cured animals within two days post-treatment. ITS1 TD PCR detected parasite recrudescence three days earlier than microscopy and had a higher positivity rate than microscopy (84.85% versus 57.58%) in 66 specimens of relapsing animals collected after treatments. Therefore, ITS1 TD PCR provides a useful tool in assessment of drug efficacy against T. congolense infection in cattle. As the assay bears the potential for detection of mixed infections, it may be applicable for drug efficacy studies and diagnostic discrimination of T. vivax and T. congolense against other pathogenic trypanosomes, including T. brucei, T. evansi and T. equiperdum. © 2014 The Authors. Published by Elsevier B.V.
Authors & Co-Authors
Tran, Thao
United Kingdom, Edinburgh
Galvmed, United Kingdom
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
Napier, Grant B.
United Kingdom, Edinburgh
Galvmed, United Kingdom
Rowan, Timothy G.
United Kingdom, Edinburgh
Galvmed, United Kingdom
Cordel, Claudia
South Africa, Bloemfontein
Clinvet International Pty Ltd.
Labuschagné, Michel
South Africa, Bloemfontein
Clinvet International Pty Ltd.
Delespaux, Vincent
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
van Reet, Nick
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
Erasmus, Heidi L.
South Africa, Bloemfontein
Clinvet International Pty Ltd.
Joubert, Annesca
South Africa, Bloemfontein
Clinvet International Pty Ltd.
Büscher, Philippe
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
Statistics
Citations: 19
Authors: 10
Affiliations: 3
Identifiers
Doi:
10.1016/j.vetpar.2014.03.005
ISSN:
03044017
Research Areas
Genetics And Genomics
Study Design
Cohort Study