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Evaluation of recombinant outer membrane protein C based indirect enzyme-linked immunoassay for the detection of Salmonella antibodies in poultry

AIM: To evaluate the efficacy of recombinant outer membrane proteinC (rOmpC) based enzyme-linked immunoassay (ELISA) for the diagnosis of salmonellosis in poultry. MATERIALS AND METHODS: Three antigens were prepared, and the indirect ELISA was standardized using the antigens and the antiserum raised...

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Autores principales: Manoj, Jinu, Agarwal, Rajesh K., Sailo, Blessa, Wani, Mudasir Ahmed, Singh, Manoj Kumar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Veterinary World 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4774754/
https://www.ncbi.nlm.nih.gov/pubmed/27047189
http://dx.doi.org/10.14202/vetworld.2015.1006-1010
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author Manoj, Jinu
Agarwal, Rajesh K.
Sailo, Blessa
Wani, Mudasir Ahmed
Singh, Manoj Kumar
author_facet Manoj, Jinu
Agarwal, Rajesh K.
Sailo, Blessa
Wani, Mudasir Ahmed
Singh, Manoj Kumar
author_sort Manoj, Jinu
collection PubMed
description AIM: To evaluate the efficacy of recombinant outer membrane proteinC (rOmpC) based enzyme-linked immunoassay (ELISA) for the diagnosis of salmonellosis in poultry. MATERIALS AND METHODS: Three antigens were prepared, and the indirect ELISA was standardized using the antigens and the antiserum raised in chicken against Omp and rOmpC. Sera were collected from a total of 255 apparently healthy field chickens and screened for the presence of Salmonella antibodies by this ELISA. RESULTS: The sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of Omp revealed major polypeptides at 36, 42 and 52 kDa, and the rOmpC was evident by a single protein band of 43 kDa. The Omp and rOmpC antigen revealed an optimum concentration of 78 and 156 ng, respectively, in the assay, while the whole cell antigen gave an optimum reaction at a concentration of 10(6) organisms/ml. The test was found to be specific as it did not react with any of the antisera of seven other organisms. The developed ELISA detected Salmonella antibodies from 22 (8.62%) samples with rOmpC antigen, while 24 (9.41%) samples gave a positive reaction with both Omp and whole cell antigens. CONCLUSION: We suggest rOmpC based indirect ELISA as a suitable screening tool for serological monitoring of poultry flocks.
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spelling pubmed-47747542016-04-04 Evaluation of recombinant outer membrane protein C based indirect enzyme-linked immunoassay for the detection of Salmonella antibodies in poultry Manoj, Jinu Agarwal, Rajesh K. Sailo, Blessa Wani, Mudasir Ahmed Singh, Manoj Kumar Vet World Research Article AIM: To evaluate the efficacy of recombinant outer membrane proteinC (rOmpC) based enzyme-linked immunoassay (ELISA) for the diagnosis of salmonellosis in poultry. MATERIALS AND METHODS: Three antigens were prepared, and the indirect ELISA was standardized using the antigens and the antiserum raised in chicken against Omp and rOmpC. Sera were collected from a total of 255 apparently healthy field chickens and screened for the presence of Salmonella antibodies by this ELISA. RESULTS: The sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of Omp revealed major polypeptides at 36, 42 and 52 kDa, and the rOmpC was evident by a single protein band of 43 kDa. The Omp and rOmpC antigen revealed an optimum concentration of 78 and 156 ng, respectively, in the assay, while the whole cell antigen gave an optimum reaction at a concentration of 10(6) organisms/ml. The test was found to be specific as it did not react with any of the antisera of seven other organisms. The developed ELISA detected Salmonella antibodies from 22 (8.62%) samples with rOmpC antigen, while 24 (9.41%) samples gave a positive reaction with both Omp and whole cell antigens. CONCLUSION: We suggest rOmpC based indirect ELISA as a suitable screening tool for serological monitoring of poultry flocks. Veterinary World 2015-08 2015-08-25 /pmc/articles/PMC4774754/ /pubmed/27047189 http://dx.doi.org/10.14202/vetworld.2015.1006-1010 Text en Copyright: © The authors. http://creativecommons.org/licenses/by/2.0 This article is an open access article licensed under the terms of the Creative Commons Attributin License (http://creative commons.org/licenses/by/2.0) which permits unrestricted use, distribution and reproduction in any medium, provided the work is properly cited.
spellingShingle Research Article
Manoj, Jinu
Agarwal, Rajesh K.
Sailo, Blessa
Wani, Mudasir Ahmed
Singh, Manoj Kumar
Evaluation of recombinant outer membrane protein C based indirect enzyme-linked immunoassay for the detection of Salmonella antibodies in poultry
title Evaluation of recombinant outer membrane protein C based indirect enzyme-linked immunoassay for the detection of Salmonella antibodies in poultry
title_full Evaluation of recombinant outer membrane protein C based indirect enzyme-linked immunoassay for the detection of Salmonella antibodies in poultry
title_fullStr Evaluation of recombinant outer membrane protein C based indirect enzyme-linked immunoassay for the detection of Salmonella antibodies in poultry
title_full_unstemmed Evaluation of recombinant outer membrane protein C based indirect enzyme-linked immunoassay for the detection of Salmonella antibodies in poultry
title_short Evaluation of recombinant outer membrane protein C based indirect enzyme-linked immunoassay for the detection of Salmonella antibodies in poultry
title_sort evaluation of recombinant outer membrane protein c based indirect enzyme-linked immunoassay for the detection of salmonella antibodies in poultry
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4774754/
https://www.ncbi.nlm.nih.gov/pubmed/27047189
http://dx.doi.org/10.14202/vetworld.2015.1006-1010
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