Development of a lion-specific interferon-gamma assay

M Maas, P J S van Kooten, J Schreuder, D Morar, E Tijhaar, A L Michel, V P M G Rutten

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

The ongoing spread of bovine tuberculosis (BTB) in African free-ranging lion populations, for example in the Kruger National Park, raises the need for diagnostic assays for BTB in lions. These, in addition, would be highly relevant for zoological gardens worldwide that want to determine the BTB status of their lions, e.g. for translocations. The present study concerns the development of a lion-specific IFN-γ assay, following the production and characterization of monoclonal antibodies specific for lion interferon-gamma (IFN-γ). Recombinant lion IFN-γ (rLIFN-γ) was produced in mammalian cells and used to immunize mice to establish hybridoma cell lines producing monoclonal antibodies. These were used to develop a sensitive, lion IFN-γ-specific capture ELISA, able to detect rLIFN-γ to the level of 160 pg/ml. Recognition of native lion IFN-γ was shown in an initial assessment of supernatants of mitogen stimulated whole blood cultures of 11 known BTB-negative lions. In conclusion, the capture ELISA shows potential as a diagnostic assay for bovine tuberculosis in lions. Preliminary results also indicate the possible use of the test for other (feline) species.

Original languageEnglish
Pages (from-to)292-7
Number of pages6
JournalVeterinary Immunology and Immunopathology
Volume149
Issue number3-4
DOIs
Publication statusPublished - 2012

Keywords

  • Animals
  • Antibodies, Monoclonal/genetics
  • Cattle
  • Enzyme-Linked Immunosorbent Assay/methods
  • Female
  • Interferon-gamma/analysis
  • Lions/blood
  • Mice
  • Mice, Inbred BALB C
  • Mycobacterium bovis/genetics
  • Recombinant Proteins/genetics
  • Sensitivity and Specificity
  • Tuberculosis/blood

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