The elephant interferon gamma assay : a contribution to diagnosis of tuberculosis in elephants

Show simple item record Angkawanish, Taweepoke Morar, Darshana Van Kooten, Peter Bontekoning, Ilona Schreuder, Janneke Maas, Miriam Wajjwalku, Worawidh Sirimalaisuwan, Anucha Michel, Anita Luise Tijhaar, Edwin Rutten, Victor P.M.G. 2014-05-21T10:24:57Z 2014-12-01T00:20:05Z 2013-11
dc.description.abstract Mycobacterium tuberculosis (M. tb) has been shown to be the main causative agent of tuberculosis in elephants worldwide. M. tb may be transmitted from infected humans to other species including elephants and vice versa, in case of prolonged intensive contact. An accurate diagnostic approach covering all phases of the infection in elephants is required. As M. tb is an intracellular pathogen and cell-mediated immune (CMI) responses are elicited early after infection, the skin test is the CMI assay of choice in humans and cattle. However, this test is not applicable in elephants. The interferon gamma (IFN-γ) assay is considered a good alternative for the skin test in general, validated for use in cattle and humans. This study was aimed at development of an IFN-γ assay applicable for diagnosis of tuberculosis in elephants. Recombinant elephant IFN-γ (rEpIFN-γ) produced in eukaryotic cells was used to immunize mice and generate the monoclonal antibodies. Hybridomas were screened for IFN- γ-specific monoclonal antibody production and subcloned, and antibodies were isotyped and affinity purified. Western blot confirmed recognition of the rEpIFN-γ. The optimal combination of capture and detection antibodies selected was able to detect rEpIFN-γ in concentrations as low as 1 pg/ml. The assay was shown to be able to detect the native elephant IFN-γ, elicited in positive-control cultures (pokeweed mitogen (PWM), phorbol myristate acetate plus ionomycin (PMA/I)) of both Asian and African elephant whole-blood cultures (WBC). Preliminary data were generated using WBC from non-infected elephants, a M. tb infection-suspected elephant and a culture-confirmed M. tb-infected elephant. The latter showed measurable production of IFN-γ after stimulation with ESAT6/CFP10 PPDB and PPDA in concentration ranges as elicited in WBC by Mycobacterium tuberculosis complex (MTBC)-specific antigens in other species. Hence, the IFN-γ assay presented potential as a diagnostic tool for the detection of elephant tuberculosis. Validation of the assay will require its application in large populations of non-infected and infected elephants. en
dc.description.librarian hb2014 en
dc.description.librarian ab2014
dc.description.sponsorship EU-Asia Link project, TH/Asia-Link/012(141-055) en
dc.description.uri en
dc.identifier.citation Angkawanish, T, Morar, D, Van Kooten, P, Bontekoning, I, Schreuder, J, Maas, M, Wajjwalku, W, Sirimalaisuwan, A, Michel, AL, Tijhaar, E & Rutten, VPMG 2013, 'The elephant interferon gamma assay : a contribution to diagnosis of tuberculosis in elephants', Transboundary and Emerging Diseases, vol. 60, Suppl.1, pp. 53-59. en
dc.identifier.issn 1865-1674 (print)
dc.identifier.issn 1865-1682 (online)
dc.identifier.other 10.1111/tbed.12098
dc.identifier.other 55978917900
dc.identifier.other N-8996-2014
dc.language.iso en en
dc.publisher Wiley-Blackwell en
dc.relation.requires Adobe Acrobat Reader en
dc.rights © 2013 Blackwell Verlag GmbH. The definite version is available at : en
dc.subject Interferon gamma en
dc.subject M. tb en
dc.subject.lcsh Elephants en
dc.subject.lcsh Mycobacterium tuberculosis en
dc.subject.lcsh Interferon en
dc.subject.lcsh Tuberculosis en
dc.title The elephant interferon gamma assay : a contribution to diagnosis of tuberculosis in elephants en
dc.type Preprint Article en

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