The major etiological agent of rabies, rabies virus (RABV), accounts for tens of thousands of human deaths per annum. The
majority of these deaths are associated with rabies cycles in dogs in resource-limited countries of Africa and Asia. Although
routine rabies diagnosis plays an integral role in disease surveillance and management, the application of the currently
recommended direct fluorescent antibody (DFA) test in countries on the African and Asian continents remains quite limited.
A novel diagnostic assay, the direct rapid immunohistochemical test (dRIT), has been reported to have a diagnostic
sensitivity and specificity equal to that of the DFA test while offering advantages in cost, time and interpretation. Prior
studies used the dRIT utilized monoclonal antibody (MAb) cocktails. The objective of this study was to test the hypothesis
that a biotinylated polyclonal antibody (PAb) preparation, applied in the dRIT protocol, would yield equal or improved
results compared to the use of dRIT with MAbs. We also wanted to compare the PAb dRIT with the DFA test, utilizing the
same PAb preparation with a fluorescent label. The PAb dRIT had a diagnostic sensitivity and specificity of 100%, which was
shown to be marginally higher than the diagnostic efficacy observed for the PAb DFA test. The classical dRIT, relying on
two-biotinylated MAbs, was applied to the same panel of samples and a reduced diagnostic sensitivity (83.50% and 90.78%
respectively) was observed. Antigenic typing of the false negative samples indicated all of these to be mongoose RABV
variants. Our results provided evidence that a dRIT with alternative antibody preparations, conjugated to a biotin moiety,
has a diagnostic efficacy equal to that of a DFA relying on the same antibody and that the antibody preparation should be
optimized for virus variants specific to the geographical area of focus.