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Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids
The high abortion rate associated with Salmonella Abortusequi (S. Abortusequi) infection in equids has re-emerged over the past 10 years and has caused serious economic losses to China. Our previous studies showed that the flagellin FljB gene could distinguish S. Abortusequi from most Salmonella ser...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10662346/ https://www.ncbi.nlm.nih.gov/pubmed/37874302 http://dx.doi.org/10.1128/jcm.00273-23 |
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author | Guo, Kui Guo, Wei Liu, Diqiu Zhang, Weiguo Yang, Yan Zhang, Zenan Li, Shuaijie Wang, Jinhui Chu, Xiaoyu Wang, Yaoxin Hu, Zhe Wang, Xiaojun |
author_facet | Guo, Kui Guo, Wei Liu, Diqiu Zhang, Weiguo Yang, Yan Zhang, Zenan Li, Shuaijie Wang, Jinhui Chu, Xiaoyu Wang, Yaoxin Hu, Zhe Wang, Xiaojun |
author_sort | Guo, Kui |
collection | PubMed |
description | The high abortion rate associated with Salmonella Abortusequi (S. Abortusequi) infection in equids has re-emerged over the past 10 years and has caused serious economic losses to China. Our previous studies showed that the flagellin FljB gene could distinguish S. Abortusequi from most Salmonella serotypes. In this study, the flagellin antigen was used to develop a competitive enzyme-linked immunosorbent assay (cELISA) that could be used to detect both horse and donkey serum samples using a monoclonal antibody (MAb) that was found to bind to FljB. A cELISA was established using the purified MAb coating of the plate and incubation of the mixture of horseradish peroxidase (HRP)-conjugated FljB antigen with the undiluted serum sample. The performance of the cELISA and the tube agglutination test (TAT) assay was compared with respect to sensitivity and specificity, by testing a panel containing 660 S. Abortusequi-positive and 515 S. Abortusequi-negative serum samples, all of which had been characterized by Western blotting. Receiver operator characteristic (ROC) analyses were performed to determine the cutoff value and estimate the detection specificity (Sp) and sensitivity (Se). ROC analysis showed that the area under the ROC curve (AUC) values of cELISA [AUC = 0.9941; 95% confidence interval (CI), 0.9898–0.9984] were higher than those of TAT (AUC = 0.7705; 95% Cl, 0.7437–0.7972). A cutoff value of 39.5% was selected with Sp and Se values of 100 (95% Cl, 99.26–100.00) and 97.58 (95% Cl, 96.10–98.50), respectively. The cELISA has excellent futures compared with TAT, such as shortened detection time, no need for pre-treatment of sera, and easy interpretation of the results, and is more suitable for disease surveillance. |
format | Online Article Text |
id | pubmed-10662346 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-106623462023-10-24 Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids Guo, Kui Guo, Wei Liu, Diqiu Zhang, Weiguo Yang, Yan Zhang, Zenan Li, Shuaijie Wang, Jinhui Chu, Xiaoyu Wang, Yaoxin Hu, Zhe Wang, Xiaojun J Clin Microbiol Clinical Veterinary Microbiology The high abortion rate associated with Salmonella Abortusequi (S. Abortusequi) infection in equids has re-emerged over the past 10 years and has caused serious economic losses to China. Our previous studies showed that the flagellin FljB gene could distinguish S. Abortusequi from most Salmonella serotypes. In this study, the flagellin antigen was used to develop a competitive enzyme-linked immunosorbent assay (cELISA) that could be used to detect both horse and donkey serum samples using a monoclonal antibody (MAb) that was found to bind to FljB. A cELISA was established using the purified MAb coating of the plate and incubation of the mixture of horseradish peroxidase (HRP)-conjugated FljB antigen with the undiluted serum sample. The performance of the cELISA and the tube agglutination test (TAT) assay was compared with respect to sensitivity and specificity, by testing a panel containing 660 S. Abortusequi-positive and 515 S. Abortusequi-negative serum samples, all of which had been characterized by Western blotting. Receiver operator characteristic (ROC) analyses were performed to determine the cutoff value and estimate the detection specificity (Sp) and sensitivity (Se). ROC analysis showed that the area under the ROC curve (AUC) values of cELISA [AUC = 0.9941; 95% confidence interval (CI), 0.9898–0.9984] were higher than those of TAT (AUC = 0.7705; 95% Cl, 0.7437–0.7972). A cutoff value of 39.5% was selected with Sp and Se values of 100 (95% Cl, 99.26–100.00) and 97.58 (95% Cl, 96.10–98.50), respectively. The cELISA has excellent futures compared with TAT, such as shortened detection time, no need for pre-treatment of sera, and easy interpretation of the results, and is more suitable for disease surveillance. American Society for Microbiology 2023-10-24 /pmc/articles/PMC10662346/ /pubmed/37874302 http://dx.doi.org/10.1128/jcm.00273-23 Text en Copyright © 2023 Guo et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Clinical Veterinary Microbiology Guo, Kui Guo, Wei Liu, Diqiu Zhang, Weiguo Yang, Yan Zhang, Zenan Li, Shuaijie Wang, Jinhui Chu, Xiaoyu Wang, Yaoxin Hu, Zhe Wang, Xiaojun Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids |
title | Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids |
title_full | Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids |
title_fullStr | Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids |
title_full_unstemmed | Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids |
title_short | Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids |
title_sort | development and application of a competitive elisa for the detection of antibodies against salmonella abortusequi in equids |
topic | Clinical Veterinary Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10662346/ https://www.ncbi.nlm.nih.gov/pubmed/37874302 http://dx.doi.org/10.1128/jcm.00273-23 |
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