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Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels
Epizootic haemorrhagic disease virus (EHDV) is a member of the Orbivirus genus in the Reoviridae family, and it is the etiological agent of an arthropod-transmitted disease that affects domestic and wild ruminants. Due to its significant economic impact, many attempts have been done in order to deve...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8959998/ https://www.ncbi.nlm.nih.gov/pubmed/35356489 http://dx.doi.org/10.1155/2022/5210771 |
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author | Serroni, Anna Ulisse, Simonetta Iorio, Mariangela Laguardia, Caterina Testa, Lilia Armillotta, Gisella Caporale, Marco Salini, Romolo Lelli, Davide Wernery, Ulrich Raghavan, Rekha Mercante, Maria Teresa Di Ventura, Mauro |
author_facet | Serroni, Anna Ulisse, Simonetta Iorio, Mariangela Laguardia, Caterina Testa, Lilia Armillotta, Gisella Caporale, Marco Salini, Romolo Lelli, Davide Wernery, Ulrich Raghavan, Rekha Mercante, Maria Teresa Di Ventura, Mauro |
author_sort | Serroni, Anna |
collection | PubMed |
description | Epizootic haemorrhagic disease virus (EHDV) is a member of the Orbivirus genus in the Reoviridae family, and it is the etiological agent of an arthropod-transmitted disease that affects domestic and wild ruminants. Due to its significant economic impact, many attempts have been done in order to develop diagnostic immunoassays mainly based on the use of the viral protein 7 (VP7), that is, the immunodominant serogroup-specific antigen. In this work, a recombinant VP7 (recVP7) of EHDV serotype 2 was produced in a baculovirus system, and after purification using ion metal affinity chromatography, we obtained a high yield of recombinant protein characterized by a high degree of purity. We used the purified recVP7 as reagent to develop a competitive enzyme-linked immunoassay (c-ELISA), and we tested the presence of EHDV antibodies in 185 dromedary camel serum samples. The c-ELISA showed good performance parameters in recognising positive sera of naturally EHDV-infected dromedary camels; in particular, our developed test reached 85.7% of sensitivity, 98.1% of specificity, 93% of accuracy, and a high agreement value with results obtained by the commercial ELISA kit (Cohen's kappa value of 0.85) that we adopted as the reference method. This c-ELISA could be a useful screening test to monitor the virus spread in camels that are sentinel animals for endemic areas of disease. |
format | Online Article Text |
id | pubmed-8959998 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
spelling | pubmed-89599982022-03-29 Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels Serroni, Anna Ulisse, Simonetta Iorio, Mariangela Laguardia, Caterina Testa, Lilia Armillotta, Gisella Caporale, Marco Salini, Romolo Lelli, Davide Wernery, Ulrich Raghavan, Rekha Mercante, Maria Teresa Di Ventura, Mauro J Trop Med Research Article Epizootic haemorrhagic disease virus (EHDV) is a member of the Orbivirus genus in the Reoviridae family, and it is the etiological agent of an arthropod-transmitted disease that affects domestic and wild ruminants. Due to its significant economic impact, many attempts have been done in order to develop diagnostic immunoassays mainly based on the use of the viral protein 7 (VP7), that is, the immunodominant serogroup-specific antigen. In this work, a recombinant VP7 (recVP7) of EHDV serotype 2 was produced in a baculovirus system, and after purification using ion metal affinity chromatography, we obtained a high yield of recombinant protein characterized by a high degree of purity. We used the purified recVP7 as reagent to develop a competitive enzyme-linked immunoassay (c-ELISA), and we tested the presence of EHDV antibodies in 185 dromedary camel serum samples. The c-ELISA showed good performance parameters in recognising positive sera of naturally EHDV-infected dromedary camels; in particular, our developed test reached 85.7% of sensitivity, 98.1% of specificity, 93% of accuracy, and a high agreement value with results obtained by the commercial ELISA kit (Cohen's kappa value of 0.85) that we adopted as the reference method. This c-ELISA could be a useful screening test to monitor the virus spread in camels that are sentinel animals for endemic areas of disease. Hindawi 2022-03-21 /pmc/articles/PMC8959998/ /pubmed/35356489 http://dx.doi.org/10.1155/2022/5210771 Text en Copyright © 2022 Anna Serroni et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Serroni, Anna Ulisse, Simonetta Iorio, Mariangela Laguardia, Caterina Testa, Lilia Armillotta, Gisella Caporale, Marco Salini, Romolo Lelli, Davide Wernery, Ulrich Raghavan, Rekha Mercante, Maria Teresa Di Ventura, Mauro Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels |
title | Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels |
title_full | Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels |
title_fullStr | Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels |
title_full_unstemmed | Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels |
title_short | Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels |
title_sort | development of a competitive enzyme-linked immunosorbent assay based on purified recombinant viral protein 7 for serological diagnosis of epizootic haemorrhagic disease in camels |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8959998/ https://www.ncbi.nlm.nih.gov/pubmed/35356489 http://dx.doi.org/10.1155/2022/5210771 |
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