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Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein
BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating diseases affecting the swine industry globally. Evaluation of antibody responses and neutralizing antibody titers is the most effective method for vaccine evaluation. In this study, the B cel...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8602981/ https://www.ncbi.nlm.nih.gov/pubmed/34798885 http://dx.doi.org/10.1186/s12917-021-03060-z |
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author | Zhao, Jun Zhang, Rubo Zhu, Ling Deng, Huidan Li, Fengqing Xu, Lei Huan, Jianbo Sun, Xiangang Xu, Zhiwen |
author_facet | Zhao, Jun Zhang, Rubo Zhu, Ling Deng, Huidan Li, Fengqing Xu, Lei Huan, Jianbo Sun, Xiangang Xu, Zhiwen |
author_sort | Zhao, Jun |
collection | PubMed |
description | BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating diseases affecting the swine industry globally. Evaluation of antibody responses and neutralizing antibody titers is the most effective method for vaccine evaluation. In this study, the B cell line epitopes of PRRSV M protein were predicted, and two peptide ELISA assays were established (M-A110-129 ELISA, M-A148-174 ELISA) to detect antibodies against PRRSV M protein. Field serum samples collected from pig farms were used to validate the peptide ELISA and compare it with an indirect immunofluorescence assay. RESULTS: The sensitivity and specificity of M-A110-129 ELISA and M-A148-174 ELISA were (111/125) 88.80%, (69/70) 98.57% and (122/125) 97.60%, (70/70) 100%, relative to indirect immunofluorescence assay. This peptide ELISA could detect antibodies against different genotypes of PRRSV including type 1 PRRSV, classical PRRSV, HP-PRRSV, and NADC30 like PRRSV, but not antibodies against other common swine viruses. The results of ROC analysis showed that the area under the curve (AUC) of the M-A110-129 ELISA and M-A148-174 ELISA were 0.967 and 0.996, respectively. Compared the concordance of results using two peptide ELISA assays, the IDEXX PRRSV X3 Ab ELISA and a virus neutralization test, were assessed using a series of 147 sera from pigs vaccinated with the NADC30-like PRRSV inactivated vaccine. The M-A148-174 ELISA had the best consistency, with a Cohen’s kappa coefficient of 0.8772. The concordance rates of the Hipra PRRSV ELISA kit, M-A110-129 ELISA and M-A148-174 ELISA in the field seropositive detection results were 91.08, 86.32 and 95.35%, relative to indirect immunofluorescence assay. CONCLUSIONS: In summary, compared with M-A110-129 ELISA, the PRRSV M-A148-174 ELISA is of value for detecting antibodies against PRRSV and the evaluation of the NADC30-like PRRSV inactivated vaccine, but the advantage is insufficient in serological early diagnosis. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12917-021-03060-z. |
format | Online Article Text |
id | pubmed-8602981 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-86029812021-11-19 Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein Zhao, Jun Zhang, Rubo Zhu, Ling Deng, Huidan Li, Fengqing Xu, Lei Huan, Jianbo Sun, Xiangang Xu, Zhiwen BMC Vet Res Research BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating diseases affecting the swine industry globally. Evaluation of antibody responses and neutralizing antibody titers is the most effective method for vaccine evaluation. In this study, the B cell line epitopes of PRRSV M protein were predicted, and two peptide ELISA assays were established (M-A110-129 ELISA, M-A148-174 ELISA) to detect antibodies against PRRSV M protein. Field serum samples collected from pig farms were used to validate the peptide ELISA and compare it with an indirect immunofluorescence assay. RESULTS: The sensitivity and specificity of M-A110-129 ELISA and M-A148-174 ELISA were (111/125) 88.80%, (69/70) 98.57% and (122/125) 97.60%, (70/70) 100%, relative to indirect immunofluorescence assay. This peptide ELISA could detect antibodies against different genotypes of PRRSV including type 1 PRRSV, classical PRRSV, HP-PRRSV, and NADC30 like PRRSV, but not antibodies against other common swine viruses. The results of ROC analysis showed that the area under the curve (AUC) of the M-A110-129 ELISA and M-A148-174 ELISA were 0.967 and 0.996, respectively. Compared the concordance of results using two peptide ELISA assays, the IDEXX PRRSV X3 Ab ELISA and a virus neutralization test, were assessed using a series of 147 sera from pigs vaccinated with the NADC30-like PRRSV inactivated vaccine. The M-A148-174 ELISA had the best consistency, with a Cohen’s kappa coefficient of 0.8772. The concordance rates of the Hipra PRRSV ELISA kit, M-A110-129 ELISA and M-A148-174 ELISA in the field seropositive detection results were 91.08, 86.32 and 95.35%, relative to indirect immunofluorescence assay. CONCLUSIONS: In summary, compared with M-A110-129 ELISA, the PRRSV M-A148-174 ELISA is of value for detecting antibodies against PRRSV and the evaluation of the NADC30-like PRRSV inactivated vaccine, but the advantage is insufficient in serological early diagnosis. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12917-021-03060-z. BioMed Central 2021-11-19 /pmc/articles/PMC8602981/ /pubmed/34798885 http://dx.doi.org/10.1186/s12917-021-03060-z Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Zhao, Jun Zhang, Rubo Zhu, Ling Deng, Huidan Li, Fengqing Xu, Lei Huan, Jianbo Sun, Xiangang Xu, Zhiwen Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein |
title | Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein |
title_full | Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein |
title_fullStr | Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein |
title_full_unstemmed | Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein |
title_short | Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein |
title_sort | establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against prrsv m protein |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8602981/ https://www.ncbi.nlm.nih.gov/pubmed/34798885 http://dx.doi.org/10.1186/s12917-021-03060-z |
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