1. Development of biotinylated polyclonal anti-ribonucleoprotein IgG for detection of rabies virus antigen by direct rapid immunohistochemical test.
- Author
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Castro BS, Guedes F, Fernandes ER, Koike G, Katz ISS, Chaves LB, and Silva SR
- Subjects
- Animals, Antibodies, Monoclonal metabolism, Antibodies, Viral immunology, Antibodies, Viral metabolism, Biotinylation, Brain immunology, Brain virology, Cats, Cattle, Chiroptera, Dogs, Fluorescent Antibody Technique, Direct methods, Horses, Immunoglobulin G metabolism, Immunohistochemistry methods, Primates, Rabies diagnosis, Rabies virology, Sensitivity and Specificity, Species Specificity, Swine, Antibodies, Monoclonal immunology, Antigens, Viral immunology, Immunoglobulin G immunology, Rabies immunology, Rabies virus immunology, Ribonucleoproteins immunology
- Abstract
The direct rapid immunohistochemical test (dRIT) has been recommended for laboratorial diagnosis of rabies, especially in developing countries. The absence of commercial primary antibodies, however, still represents a major limitation to its wider use in testing. We describe here the development of a biotinylated polyclonal antibody against Rabies lyssavirus (RABV) ribonucleoprotein (RNP) and its use as a primary reagent in dRIT. Anti-RNP polyclonal horse IgG was purified by ionic exchange chromatography followed by immunoaffinity column chromatography, and its affinity, diagnostic sensitivity, and specificity were evaluated. CNS samples (120) of suspected rabies cases in different animal species were tested by dRIT, with the positive (n = 14) and negative (n = 106) results confirmed by direct fluorescence antibody testing (dFAT). Comparing the results of dRIT and dFAT, we found that the biotinylated anti-RNP IgG delivered 100% diagnostic specificity and sensibility for rabies diagnosis. Our findings show that the biotinylated anti-RNP polyclonal IgG can be produced with the quality required for application in dRIT. This work represents an important step in efforts to diagnose rabies in developing countries., (Copyright © 2020 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2020
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