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Development of an effective one-step double-antigen sandwich ELISA based on p72 to detect antibodies against African swine fever virus
African swine fever (ASF), caused by ASF virus (ASFV), is a highly contagious and lethal disease of domestic pigs leading to tremendous economic losses. As there are no vaccines and drugs available. An effective diagnosis to eliminate ASFV-infected pigs is a crucial strategy to prevent and control A...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10287978/ https://www.ncbi.nlm.nih.gov/pubmed/37360404 http://dx.doi.org/10.3389/fvets.2023.1160583 |
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author | Wang, Lei Li, Duan Liu, Yanlin Zhang, Leyi Peng, Guoliang Xu, Zheng Jia, Hong Song, Changxu |
author_facet | Wang, Lei Li, Duan Liu, Yanlin Zhang, Leyi Peng, Guoliang Xu, Zheng Jia, Hong Song, Changxu |
author_sort | Wang, Lei |
collection | PubMed |
description | African swine fever (ASF), caused by ASF virus (ASFV), is a highly contagious and lethal disease of domestic pigs leading to tremendous economic losses. As there are no vaccines and drugs available. An effective diagnosis to eliminate ASFV-infected pigs is a crucial strategy to prevent and control ASF. To this end, ASFV capsid protein p72 was expressed using Chinese hamster ovary (CHO) cells and subsequently conjugated with horseradish peroxidase (HRP) to develop a one-step double-antigen sandwich enzyme-linked immunosorbent assay (one-step DAgS-ELISA). The performance of this ELISA for detecting ASFV antibodies was evaluated. Overall, a diagnostic sensitivity of 97.96% and specificity of 98.96% was achieved when the cutoff value was set to 0.25. No cross-reaction with healthy pig serum and other swine viruses was observed. The coefficients of variation of the intra-assay and inter-assay were both <10%. Importantly, this ELISA could detect antibodies in standard serum with 12,800-fold dilution, and seroconversion started from the 7th day post-inoculation (dpi), showing excellent analytical sensitivity and great utility. Furthermore, compared to the commercial kit, this ELISA had a good agreement and significantly shorter operation time. Collectively, a novel one-step DAgS-ELISA for detecting antibodies against ASFV is developed, which will be reliable and convenient to monitor ASFV infection. |
format | Online Article Text |
id | pubmed-10287978 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-102879782023-06-24 Development of an effective one-step double-antigen sandwich ELISA based on p72 to detect antibodies against African swine fever virus Wang, Lei Li, Duan Liu, Yanlin Zhang, Leyi Peng, Guoliang Xu, Zheng Jia, Hong Song, Changxu Front Vet Sci Veterinary Science African swine fever (ASF), caused by ASF virus (ASFV), is a highly contagious and lethal disease of domestic pigs leading to tremendous economic losses. As there are no vaccines and drugs available. An effective diagnosis to eliminate ASFV-infected pigs is a crucial strategy to prevent and control ASF. To this end, ASFV capsid protein p72 was expressed using Chinese hamster ovary (CHO) cells and subsequently conjugated with horseradish peroxidase (HRP) to develop a one-step double-antigen sandwich enzyme-linked immunosorbent assay (one-step DAgS-ELISA). The performance of this ELISA for detecting ASFV antibodies was evaluated. Overall, a diagnostic sensitivity of 97.96% and specificity of 98.96% was achieved when the cutoff value was set to 0.25. No cross-reaction with healthy pig serum and other swine viruses was observed. The coefficients of variation of the intra-assay and inter-assay were both <10%. Importantly, this ELISA could detect antibodies in standard serum with 12,800-fold dilution, and seroconversion started from the 7th day post-inoculation (dpi), showing excellent analytical sensitivity and great utility. Furthermore, compared to the commercial kit, this ELISA had a good agreement and significantly shorter operation time. Collectively, a novel one-step DAgS-ELISA for detecting antibodies against ASFV is developed, which will be reliable and convenient to monitor ASFV infection. Frontiers Media S.A. 2023-06-09 /pmc/articles/PMC10287978/ /pubmed/37360404 http://dx.doi.org/10.3389/fvets.2023.1160583 Text en Copyright © 2023 Wang, Li, Liu, Zhang, Peng, Xu, Jia and Song. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Veterinary Science Wang, Lei Li, Duan Liu, Yanlin Zhang, Leyi Peng, Guoliang Xu, Zheng Jia, Hong Song, Changxu Development of an effective one-step double-antigen sandwich ELISA based on p72 to detect antibodies against African swine fever virus |
title | Development of an effective one-step double-antigen sandwich ELISA based on p72 to detect antibodies against African swine fever virus |
title_full | Development of an effective one-step double-antigen sandwich ELISA based on p72 to detect antibodies against African swine fever virus |
title_fullStr | Development of an effective one-step double-antigen sandwich ELISA based on p72 to detect antibodies against African swine fever virus |
title_full_unstemmed | Development of an effective one-step double-antigen sandwich ELISA based on p72 to detect antibodies against African swine fever virus |
title_short | Development of an effective one-step double-antigen sandwich ELISA based on p72 to detect antibodies against African swine fever virus |
title_sort | development of an effective one-step double-antigen sandwich elisa based on p72 to detect antibodies against african swine fever virus |
topic | Veterinary Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10287978/ https://www.ncbi.nlm.nih.gov/pubmed/37360404 http://dx.doi.org/10.3389/fvets.2023.1160583 |
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