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Development of an Indirect ELISA to Detect African Swine Fever Virus pp62 Protein-Specific Antibodies
African swine fever (ASF) is a highly detrimental viral disease caused by African swine fever virus (ASFV). The occurrence and prevalence of this disease have become a serious threat to the global swine industry and national economies. At present, the detection volume of African swine fever is huge,...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8792508/ https://www.ncbi.nlm.nih.gov/pubmed/35097046 http://dx.doi.org/10.3389/fvets.2021.798559 |
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author | Zhong, Kexin Zhu, Mengmeng Yuan, Qichao Deng, Zhibang Feng, Simeng Liu, Daoxin Yuan, Xiaomin |
author_facet | Zhong, Kexin Zhu, Mengmeng Yuan, Qichao Deng, Zhibang Feng, Simeng Liu, Daoxin Yuan, Xiaomin |
author_sort | Zhong, Kexin |
collection | PubMed |
description | African swine fever (ASF) is a highly detrimental viral disease caused by African swine fever virus (ASFV). The occurrence and prevalence of this disease have become a serious threat to the global swine industry and national economies. At present, the detection volume of African swine fever is huge, more sensitive and accurate detection techniques are needed for the market. pp62 protein, as a protein in the late stage of infection, has strong antigenicity and a high corresponding antibody titer in infected pigs. In this study, the CP530R gene was cloned into expression vector pET-28a to construct a prokaryotic expression plasmid, which was induced by IPTG to express soluble pp62 protein. Western blot analysis showed that it had great reactivity. Using the purified recombinant protein as an antigen, an indirect ELISA method for detecting ASFV antibody was established. The method was specific only to ASFV-positive serum, 1:1600 diluted positive serum could still be detected, and the coefficients of variation (CV) of the intra assay and inter assay were both <10%. It turns out that the assays had excellent specificity, sensitivity, and repeatability. This provides an accurate, rapid, and economical method for the detection of ASFV antibody in clinical pig serum samples. |
format | Online Article Text |
id | pubmed-8792508 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-87925082022-01-28 Development of an Indirect ELISA to Detect African Swine Fever Virus pp62 Protein-Specific Antibodies Zhong, Kexin Zhu, Mengmeng Yuan, Qichao Deng, Zhibang Feng, Simeng Liu, Daoxin Yuan, Xiaomin Front Vet Sci Veterinary Science African swine fever (ASF) is a highly detrimental viral disease caused by African swine fever virus (ASFV). The occurrence and prevalence of this disease have become a serious threat to the global swine industry and national economies. At present, the detection volume of African swine fever is huge, more sensitive and accurate detection techniques are needed for the market. pp62 protein, as a protein in the late stage of infection, has strong antigenicity and a high corresponding antibody titer in infected pigs. In this study, the CP530R gene was cloned into expression vector pET-28a to construct a prokaryotic expression plasmid, which was induced by IPTG to express soluble pp62 protein. Western blot analysis showed that it had great reactivity. Using the purified recombinant protein as an antigen, an indirect ELISA method for detecting ASFV antibody was established. The method was specific only to ASFV-positive serum, 1:1600 diluted positive serum could still be detected, and the coefficients of variation (CV) of the intra assay and inter assay were both <10%. It turns out that the assays had excellent specificity, sensitivity, and repeatability. This provides an accurate, rapid, and economical method for the detection of ASFV antibody in clinical pig serum samples. Frontiers Media S.A. 2022-01-13 /pmc/articles/PMC8792508/ /pubmed/35097046 http://dx.doi.org/10.3389/fvets.2021.798559 Text en Copyright © 2022 Zhong, Zhu, Yuan, Deng, Feng, Liu and Yuan. 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 Zhong, Kexin Zhu, Mengmeng Yuan, Qichao Deng, Zhibang Feng, Simeng Liu, Daoxin Yuan, Xiaomin Development of an Indirect ELISA to Detect African Swine Fever Virus pp62 Protein-Specific Antibodies |
title | Development of an Indirect ELISA to Detect African Swine Fever Virus pp62 Protein-Specific Antibodies |
title_full | Development of an Indirect ELISA to Detect African Swine Fever Virus pp62 Protein-Specific Antibodies |
title_fullStr | Development of an Indirect ELISA to Detect African Swine Fever Virus pp62 Protein-Specific Antibodies |
title_full_unstemmed | Development of an Indirect ELISA to Detect African Swine Fever Virus pp62 Protein-Specific Antibodies |
title_short | Development of an Indirect ELISA to Detect African Swine Fever Virus pp62 Protein-Specific Antibodies |
title_sort | development of an indirect elisa to detect african swine fever virus pp62 protein-specific antibodies |
topic | Veterinary Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8792508/ https://www.ncbi.nlm.nih.gov/pubmed/35097046 http://dx.doi.org/10.3389/fvets.2021.798559 |
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