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...
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2022
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ftdoajarticles:oai:doaj.org/article:7275bb2d317c49539e5ae6f4422ba1c3 2023-05-15T15:09:37+02:00 Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels Anna Serroni Simonetta Ulisse Mariangela Iorio Caterina Laguardia Lilia Testa Gisella Armillotta Marco Caporale Romolo Salini Davide Lelli Ulrich Wernery Rekha Raghavan Maria Teresa Mercante Mauro Di Ventura 2022-01-01T00:00:00Z https://doi.org/10.1155/2022/5210771 https://doaj.org/article/7275bb2d317c49539e5ae6f4422ba1c3 EN eng Hindawi Limited http://dx.doi.org/10.1155/2022/5210771 https://doaj.org/toc/1687-9694 1687-9694 doi:10.1155/2022/5210771 https://doaj.org/article/7275bb2d317c49539e5ae6f4422ba1c3 Journal of Tropical Medicine, Vol 2022 (2022) Arctic medicine. Tropical medicine RC955-962 article 2022 ftdoajarticles https://doi.org/10.1155/2022/5210771 2022-12-31T08:40:21Z 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. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Dromedary ENVELOPE(163.033,163.033,-78.317,-78.317) Journal of Tropical Medicine 2022 1 10 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Anna Serroni Simonetta Ulisse Mariangela Iorio Caterina Laguardia Lilia Testa Gisella Armillotta Marco Caporale Romolo Salini Davide Lelli Ulrich Wernery Rekha Raghavan Maria Teresa Mercante Mauro Di Ventura 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_facet |
Arctic medicine. Tropical medicine RC955-962 |
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 |
Article in Journal/Newspaper |
author |
Anna Serroni Simonetta Ulisse Mariangela Iorio Caterina Laguardia Lilia Testa Gisella Armillotta Marco Caporale Romolo Salini Davide Lelli Ulrich Wernery Rekha Raghavan Maria Teresa Mercante Mauro Di Ventura |
author_facet |
Anna Serroni Simonetta Ulisse Mariangela Iorio Caterina Laguardia Lilia Testa Gisella Armillotta Marco Caporale Romolo Salini Davide Lelli Ulrich Wernery Rekha Raghavan Maria Teresa Mercante Mauro Di Ventura |
author_sort |
Anna Serroni |
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_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_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_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 |
publisher |
Hindawi Limited |
publishDate |
2022 |
url |
https://doi.org/10.1155/2022/5210771 https://doaj.org/article/7275bb2d317c49539e5ae6f4422ba1c3 |
long_lat |
ENVELOPE(163.033,163.033,-78.317,-78.317) |
geographic |
Arctic Dromedary |
geographic_facet |
Arctic Dromedary |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Journal of Tropical Medicine, Vol 2022 (2022) |
op_relation |
http://dx.doi.org/10.1155/2022/5210771 https://doaj.org/toc/1687-9694 1687-9694 doi:10.1155/2022/5210771 https://doaj.org/article/7275bb2d317c49539e5ae6f4422ba1c3 |
op_doi |
https://doi.org/10.1155/2022/5210771 |
container_title |
Journal of Tropical Medicine |
container_volume |
2022 |
container_start_page |
1 |
op_container_end_page |
10 |
_version_ |
1766340774533791744 |