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Generation of mAbs to foot–and–mouth disease virus serotype A and application in a competitive ELISA for serodiagnosis
BACKGROUND: Foot–and–mouth disease (FMD) is an economically devastating disease that severely limits international trade of animals. Of the seven FMD virus (FMDV) serotypes, serotype A is one of the most widespread cross the world. Currently antibodies to FMDV are detected in animals using the virus...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5126828/ https://www.ncbi.nlm.nih.gov/pubmed/27894355 http://dx.doi.org/10.1186/s12985-016-0650-z |
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author | Yang, Ming Xu, Wanhong Bittner, Hilary Horsington, Jacquelyn Vosloo, Wilna Goolia, Melissa Lusansky, Diana Nfon, Charles |
author_facet | Yang, Ming Xu, Wanhong Bittner, Hilary Horsington, Jacquelyn Vosloo, Wilna Goolia, Melissa Lusansky, Diana Nfon, Charles |
author_sort | Yang, Ming |
collection | PubMed |
description | BACKGROUND: Foot–and–mouth disease (FMD) is an economically devastating disease that severely limits international trade of animals. Of the seven FMD virus (FMDV) serotypes, serotype A is one of the most widespread cross the world. Currently antibodies to FMDV are detected in animals using the virus neutralization test (VNT) and the enzyme-linked immunosorbent assay (ELISA). The VNT is laborious, time–consuming and reliant on live virus and cell cultures, while ELISA has the advantage of using inactivated antigens and often provides more reproducible results. The aim of this study was to develop a reliable and rapid competitive ELISA (cELISA) for the detection of antibodies to FMDV serotype A (FMDV/A). RESULTS: A panel of FMDV/A specific monoclonal antibodies (mAbs) was generated and their ability to compete with a polyclonal serum from FMDV/A–infected cattle was examined. Two mAbs inhibited the binding of a polyclonal serum to FMDV/A viruses. The binding epitopes of each were determined as conformational and located on the VP2 viral capsid protein. The FMDV/A cELISA was developed using these two mAbs and FMDV/A inactivated virus as antigen. The diagnostic specificity and sensitivity were 99.7 and 99.3% (98.5–100%) respectively, based on a predetermined cut–off of 50% inhibition. When analysing sera from animals experimentally infected with FMDV/A, the cELISA detected antibodies from 5-days post infection (dpi) and remained positive for at least 21–28 days post infection. Comparison based on the Kappa coefficient showed strong agreement (90–94%) between cELISA and VNT. CONCLUSION: The cELISA results are comparable to the VNT for antibody detection making it a simple and reliable test to detect antibodies against FMDV/A. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12985-016-0650-z) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5126828 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-51268282016-12-08 Generation of mAbs to foot–and–mouth disease virus serotype A and application in a competitive ELISA for serodiagnosis Yang, Ming Xu, Wanhong Bittner, Hilary Horsington, Jacquelyn Vosloo, Wilna Goolia, Melissa Lusansky, Diana Nfon, Charles Virol J Research BACKGROUND: Foot–and–mouth disease (FMD) is an economically devastating disease that severely limits international trade of animals. Of the seven FMD virus (FMDV) serotypes, serotype A is one of the most widespread cross the world. Currently antibodies to FMDV are detected in animals using the virus neutralization test (VNT) and the enzyme-linked immunosorbent assay (ELISA). The VNT is laborious, time–consuming and reliant on live virus and cell cultures, while ELISA has the advantage of using inactivated antigens and often provides more reproducible results. The aim of this study was to develop a reliable and rapid competitive ELISA (cELISA) for the detection of antibodies to FMDV serotype A (FMDV/A). RESULTS: A panel of FMDV/A specific monoclonal antibodies (mAbs) was generated and their ability to compete with a polyclonal serum from FMDV/A–infected cattle was examined. Two mAbs inhibited the binding of a polyclonal serum to FMDV/A viruses. The binding epitopes of each were determined as conformational and located on the VP2 viral capsid protein. The FMDV/A cELISA was developed using these two mAbs and FMDV/A inactivated virus as antigen. The diagnostic specificity and sensitivity were 99.7 and 99.3% (98.5–100%) respectively, based on a predetermined cut–off of 50% inhibition. When analysing sera from animals experimentally infected with FMDV/A, the cELISA detected antibodies from 5-days post infection (dpi) and remained positive for at least 21–28 days post infection. Comparison based on the Kappa coefficient showed strong agreement (90–94%) between cELISA and VNT. CONCLUSION: The cELISA results are comparable to the VNT for antibody detection making it a simple and reliable test to detect antibodies against FMDV/A. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12985-016-0650-z) contains supplementary material, which is available to authorized users. BioMed Central 2016-11-28 /pmc/articles/PMC5126828/ /pubmed/27894355 http://dx.doi.org/10.1186/s12985-016-0650-z Text en © The Author(s). 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Yang, Ming Xu, Wanhong Bittner, Hilary Horsington, Jacquelyn Vosloo, Wilna Goolia, Melissa Lusansky, Diana Nfon, Charles Generation of mAbs to foot–and–mouth disease virus serotype A and application in a competitive ELISA for serodiagnosis |
title | Generation of mAbs to foot–and–mouth disease virus serotype A and application in a competitive ELISA for serodiagnosis |
title_full | Generation of mAbs to foot–and–mouth disease virus serotype A and application in a competitive ELISA for serodiagnosis |
title_fullStr | Generation of mAbs to foot–and–mouth disease virus serotype A and application in a competitive ELISA for serodiagnosis |
title_full_unstemmed | Generation of mAbs to foot–and–mouth disease virus serotype A and application in a competitive ELISA for serodiagnosis |
title_short | Generation of mAbs to foot–and–mouth disease virus serotype A and application in a competitive ELISA for serodiagnosis |
title_sort | generation of mabs to foot–and–mouth disease virus serotype a and application in a competitive elisa for serodiagnosis |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5126828/ https://www.ncbi.nlm.nih.gov/pubmed/27894355 http://dx.doi.org/10.1186/s12985-016-0650-z |
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