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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...

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Autores principales: Wang, Lei, Li, Duan, Liu, Yanlin, Zhang, Leyi, Peng, Guoliang, Xu, Zheng, Jia, Hong, Song, Changxu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
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.
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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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