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Artículo

Optimization and validation of a blocking ELISA for quantitation of anti-rabies immunoglobulins in multispecies sera

Fontana, Diego SebastianIcon ; Rodriguez, María CelesteIcon ; Garay, Ernesto SergioIcon ; Russo, Susana; Prieto, ClaudioIcon
Fecha de publicación: 03/2020
Editorial: Springer
Revista: Applied Microbiology and Biotechnology
ISSN: 0175-7598
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Biotecnología relacionada con la Salud

Resumen

We developed a fast, rabies virus-free, in vitro method, based on a blocking ELISA (bELISA), to detect and accurately quantify anti-rabies glycoprotein antibodies in serum of several animal species. In this method, purified rabies virus-like particles (VLPs) are used as antigen to coat the plates, while the presence of specific rabies immunoglobulins is revealed through blocking the recognition of these VLPs by a biotinylated monoclonal antibody. A quality by design approach was carried out in order to optimize the method performance, improving the sensitivity and, thereby, reducing the limit of detection of this assay. After the method validation, we confirmed that the bELISA method is able to detect a concentration of 0.06 IU/mL rabies immunoglobulins, titer lower than the 0.5 IU/mL cutoff value established as indication for correct vaccination. Further, we assessed the correlation between bELISA, the MNT, and the Platelia methods, confirming the accuracy of this new assay. On the other hand, precision was evaluated, obtaining acceptable repeatability and intermediate precision values, showing that this bELISA could be proposed as a potential alternative method, replacing the gold standard techniques in vaccination schemes and becoming a routine control technique within regional rabies surveillance programs.
Palabras clave: BLOCKING ELISA , DESIGN OF EXPERIMENTS (DOE) , MULTISPECIES , RABIES SURVEILLANCE , VALIDATION
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info:eu-repo/semantics/restrictedAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)
Identificadores
URI: http://hdl.handle.net/11336/174647
URL: http://link.springer.com/10.1007/s00253-020-10490-6
DOI: https://doi.org/10.1007/s00253-020-10490-6
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Articulos(CCT - SANTA FE)
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - SANTA FE
Citación
Fontana, Diego Sebastian; Rodriguez, María Celeste; Garay, Ernesto Sergio; Russo, Susana; Prieto, Claudio; Optimization and validation of a blocking ELISA for quantitation of anti-rabies immunoglobulins in multispecies sera; Springer; Applied Microbiology and Biotechnology; 104; 9; 3-2020; 4127-4139
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