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Background: Feline panleukopenia virus (FPV) is an etiologic pathogen of feline panleukopenia that infects all members of Felidae including tigers (Panthera tigris). Vaccinations against FPV among wild felid species have long been practiced in zoos worldwide. However, few studies have assessed the tiger immune response post-vaccination due to the absence of a serological diagnostic tool. To address these limitations, this study aimed to develop an in-house indirect enzyme-linked immunosorbent assay (ELISA) for the monitoring of tiger antibody levels against the feline panleukopenia vaccine by employing the synthesized subunit capsid protein VP2. An in-house horseradish peroxidase (HRP) conjugated rabbit anti-tiger immunoglobulin G (IgG) polyclonal antibody (HRP-anti-tiger IgG) was produced in this study and employed in the assay. It was then compared to a commercial HRP-conjugated goat anti-cat IgG (HRP-anti-cat IgG). Sensitivity and specificity were evaluated using the Bayesian model with preferential conditional dependence between HRP-conjugated antibody-based ELISAs and hemagglutination-inhibition (HI) tests. Results: The posterior estimates for sensitivity and specificity of two indirect ELISA HRP-conjugated antibodies were higher than those of the HI test. The sensitivity and specificity of the indirect ELISA for HRP-anti-tiger IgG and HRP-anti-cat IgG were 86.5, 57.2 and 86.7%, 64.6%, respectively, while the results of the HI test were 79.1 and 54.1%. In applications, 89.6% (198/221) and 89.1% (197/221) of the tiger serum samples were determined to be seropositive by indirect ELISA testing against HRP-anti-tiger and HRP-anti-cat, respectively. Conclusion: To the best of our knowledge, the specific serology assays for the detection of the tiger IgG antibody have not yet been established. The HRP-anti-tiger IgG has been produced for the purpose of developing the specific immunoassays for tigers. Remarkably, an in-house indirect ELISA based on VP2 subunit antigen has been successfully developed in this study, providing a potentially valuable serological tool for the effective detection of tiger antibodies. [ABSTRACT FROM AUTHOR]

The immunoglobulin G (IgG) antibody is important for detecting and evaluating immune responses. Despite increasing opportunities for tigers to be exposed to emerging diseases due to global changes coupled with increasing interaction with humans, there is no available antibody specific to tigers. Aims of this study were to produce a rabbit anti-Bengal tiger (Panthera tigris tigris) IgG polyclonal antibody and to determine its specificity. The molecular weight of the purified Bengal tiger IgG was approximately 170 kDa. The cross-reactivity of rabbit anti-Bengal tiger IgG to IgG of other wild felid species and other domestic animals was analyzed by indirect enzyme-linked immunosorbent assay. The cross reactivity to other related wild animals ranged from 53.69% to 75.63%. There was high cross-reactivity with domestic cats (63.58%) and pigs (38.45%) and low cross-reactivity with domestic cows (10.46%), goats (8.60%), sheep (7.25%), and chickens (6.91%). To confirm cross-reactivity with domestic animals' IgG, western blotting was performed. These results indicated that the purified rabbit anti-Bengal tiger IgG polyclonal antibody strongly reacted with species in the family Felidae. The polyclonal antibody generated in this study has the potential to aid in the development of useful tools for further novel immunological investigations on tigers and related felid species. [ABSTRACT FROM AUTHOR]