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The Thai Journal of Veterinary Medicine

Abstract

To date, the interferon gamma (IFN-γ) assay has become an important diagnostic tool which is applied alternatively to define infection of intracellular pathogens. In this study, the molecular characteristics of IFN-γ in Rusa deer (Rusa timorensis), Hog deer (Axis porcinus), Sambar deer (Rusa unicolor) and Eld’s deer (Rucervus eldii) were determined. Among the cervid family, the analysis of IFN-γ cDNA sequences indicated 5 polymorphism nucleotide positions. Deduced IFN-γ amino acid compositions among the cervid group were negligibly different (98-100% homology). Preliminary quantification of IFN-γ mRNA expression in the cervidae by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was demonstrated. qPCR primers were designed and tested for their ability to quantify IFN-γ mRNA with a suggested detection limit at 103 target molecules. Intra- and inter-assay variations of RT-qPCR for IFN-γ were 1.41 and 4.57, respectively. Expression ratio of IFN-γ mRNA in mitogen-stimulated peripheral blood mononuclear cells (PBMCs) relative to non-stimulated cells after normalization against β2-microglobulin (β2M) in the Rusa deer, Eld’s deer and Hog deer were 6.47±3.39, 22.91±7.21 and 17.66±1.30, respectively. This preliminary demonstration may serve as the basic instrument for further development of rapid, simple and reliable immunodiagnostic methods for the study of cervidae in Thailand.

Publisher

Faculty of Veterinary Science, Chulalongkorn University

First Page

117

Last Page

122

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