Comparison of Biotinylated Monoclonal and Polyclonal Antibodies in an Evaluation of a Direct Rapid Immunohistochemical Test for the Routine Diagnosis of Rabies in Southern Africa
PLoS Neglected Tropical Diseases
Department of Microbiology and Plant Pathology, University of Pretoria, Gauteng, South Africa; Agricultural Research Council-Onderstepoort Veterinary Institute, Rabies Division, Gauteng, South Africa; Ross University School of Veterinary Medicine, Basseterre, Saint Kitts and Nevis
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. © 2014 Coetzer et al.
monoclonal antibody; polyclonal antibody; virus RNA; biotin; diagnostic kit; monoclonal antibody; virus antibody; animal tissue; Article; biotinylation; cat; cattle; central nervous system; data analysis; diagnostic test accuracy study; dog; false positive result; fluorescent antibody technique; fox; human; human tissue; immunohistochemistry; immunoreactivity; jackal; mongoose; nonhuman; nucleotide sequence; phylogeny; rabies; sensitivity and specificity; South Africa; Africa; animal; animal disease; bovine; cat disease; cattle disease; chemistry; comparative study; diagnostic kit; dog disease; immunology; isolation and purification; nervous tissue; procedures; rabies; Rabies virus; veterinary; virology; Africa, Southern; Animal Diseases; Animals; Antibodies, Monoclonal; Antibodies, Viral; Biotin; Cat Diseases; Cats; Cattle; Cattle Diseases; Dog Diseases; Dogs; Fluorescent Antibody Technique, Direct; Nerve Tissue; Rabies; Rabies virus; Reagent Kits, Diagnostic