Title page for ETD etd-03092005-080343

Document Type

Master's Dissertation

Author

Morar, Darshana

URN

etd-03092005-080343

Document Title

The development of an interferon-gamma (IFNg) assay for the diagnosis of tuberculosis caused by Mycobacterium bovis in African elephants Loxodonta Africana) and black rhinoceros (Diceros bicornis)

Degree

MSc (Veterinary Science)

Department

Veterinary Tropical Diseases

Supervisor

Advisor Name	Title
Dr A L Michel	Co-Supervisor
Dr E Tijhaar	Co-Supervisor
Dr V P M G Rutten	Co-Supervisor
Prof E Venter	Supervisor

Keywords

no key words available

Date

2003-09-10

Availability

unrestricted

Abstract

The objective of this project was to design tools for a diagnostic test that will prove valuable in the detection of tuberculosis in elephants and rhinoceros by using the cytokine IFNg as an indicator of Mycobacterium bovis responsiveness.
Interferon-gamma (IFNg), a type II interferon, is a cytokine mainly produced by Th1 and cytotoxic T-cells expressing surface markers CD4 and CD8, respectively and natural killer cells (Ibelgaufts; 1999). In response to a mycobacterial infection antigen specific Th1- and cytotoxic T-cells are induced. When these cells encounter their specific mycobacterial antigen, they will respond by producing IFNg. Based on this principle a diagnostic test was developed. In this test PBMCs will be stimulated with M.bovis specific antigen and the subsequent production of IFNg by specific T-helper cells will be determined by IFNg of elephants and rhinoceros.
In order to develop such an assay recombinant elephant and rhinoceros IFNg was cloned, sequenced, expressed, purified and subsequently a monoclonal antibody against IFNg was produced. Monoclonal antibodies were selected by a number of ELISAs using recombinant IFNg. Preliminary results are promising and further tests are underway regarding the specificity and sensitivity of the assay before field trials can be performed.
The results of this study has significant implications in the design of an IFNg diagnostic kit for the diagnosis of tuberculosis, as caused by M.bovis, in elephants and rhinoceros as well as other wildlife affected by this debilitating disease.

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