CYTOKINE-RELEASE ASSAY FOR THE DETECTION OFINFECTION IN CHEETAH ().

Abstract

The lack of species-specific assays for the diagnosis of infectious diseases, such as bovine tuberculosis, poses a threat to the management of wildlife populations, especially for vulnerable species such as cheetah (). The aim of this study was to identify and develop a cell-mediated immunological cytokine-release assay that could distinguish between-infected and uninfected cheetahs using commercially available feline cytokine ELISA and domestic cat () recombinant proteins. Antibodies against domestic cat cytokines, tumour necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), and interferon gamma (IFN-γ), were screened for cross-reactivity with plasma cytokines from cheetah whole blood stimulated using QuantiFERON-TB Gold Plus (QFT) tubes. Evidence of cytokine production in response to QFT mitogen stimulation was observed in all four ELISA assays. However only the Mabtech Cat IFN-γ ELISAkit could distinguish between-infected (= 1) and uninfected (= 1) cheetahs and was therefore selected for further evaluation. A preliminary cheetah specific cutoff value (11 pg/ml) for detectinginfection using the Mabtech Cat IFN-γ release assay was calculated using auninfected cheetah cohort. Although this study only included one confirmedculture-positive and oneculture-negative cheetah, the Mabtech Cat IFN-γ release assay demonstrated its potential for diagnostic application in this species.