Development and evaluation of an interferon-γ release assay in Asian elephants (Elephas maximus)

亚洲象(Elephas maximus)中干扰素-γ释放试验的开发和评估

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作者:Sarad Paudel, Marvin A Villanueva, Susan K Mikota, Chie Nakajima, Kamal P Gairhe, Suraj Subedi, Nabin Rayamajhi, Mariko Sashika, Michito Shimozuru, Takashi Matsuba, Yasuhiko Suzuki, Toshio Tsubota

Abstract

We developed an interferon-γ release assay (IGRA) specific for Asian elephants (Elephas maximus). Whole blood collected from forty captive Asian elephants was stimulated with three different mitogens i.e., phytohemagglutinin (PHA), pokweed mitogen (PWM) and phorbol myristate aceteate/ionomycin (PMA/I). A sandwich ELISA that was able to recognize the recombinant elephant interferon-γ (rEIFN-γ) as well as native interferon-γ from the Asian elephants was performed using anti-elephant IFN-γ rabbit polyclonal antibodies as capture antibodies and biotinylated anti-elephant IFN-γ rabbit polyclonal antibodies as detection antibodies. PMA/I was the best mitogen to use as a positive control for an Asian elephant IGRA. The development of an Asian elephant-specific IGRA that detects native IFN-γ in elephant whole blood provides promising results for its application as a potential diagnostic tool for diseases, such as tuberculosis (TB) in Asian elephants.

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