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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,...

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Autores principales: Zhong, Kexin, Zhu, Mengmeng, Yuan, Qichao, Deng, Zhibang, Feng, Simeng, Liu, Daoxin, Yuan, Xiaomin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
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.
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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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