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A commercially available Rapid Immunodiagnostic Test was evaluated using 115 brain samples collected from rabies suspected dogs, cats and cattle to determine its sensitivity and specificity under field conditions in Ethiopia. The sensitivity and specificity of the Rapid Immunodiagnostic Test Kit was assessed in comparison with the Fluorescent Antibody Test (gold standard test for rabies diagnosis). Compared to the Fluorescent Antibody Test the sensitivity and specificity of the Rapid Immunodiagnostic Test was found to be 96.5% (95% CI: 90.0–99.3%) and 100% (95% CI: 88.4– 100%), respectively. It was shown that the Rapid Immunodiagnostic Test can be used in rabies surveillance programs and in targeted post-exposure prophylaxis where the standard rabies diagnostic test is not available. The improved intensity of the result in sample stored in deep freeze shows that the Rapid Immunodiagnostic Test procedures need to be standardized. The kit should be evaluated at larger scale using samples from different species of animals at the field level.