A direct high-throughput in Cell-ELISA for measuring infectivity of cytopathic and non-cytopathic bovine viral diarrhoea virus strains applied to the assessment of antiviral activity
Low-cost high-throughput methods applicable to any virus strain are required for screening antiviral compounds against multiple field strains. Colorimetric cell-viability assays are used for this purpose as long as the viruses are cytopathic (CP) in cell culture. However, bovine viral diarrhoea viru...
| Autores principales: | , , , , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
Elsevier
2019
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| Materias: | |
| Acceso en línea: | https://www.sciencedirect.com/science/article/pii/S0166093418301617 http://hdl.handle.net/20.500.12123/6503 https://doi.org/10.1016/j.jviromet.2018.07.010 |
| _version_ | 1855035688451309568 |
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| author | Quintana, Maria Eugenia Barone, Lucas Forlenza, María Belén Trotta, Myrian Vanesa Turco, Cecilia Mansilla, Florencia Celeste Cardoso, Nancy Patricia Capozzo, Alejandra Victoria |
| author_browse | Barone, Lucas Capozzo, Alejandra Victoria Cardoso, Nancy Patricia Forlenza, María Belén Mansilla, Florencia Celeste Quintana, Maria Eugenia Trotta, Myrian Vanesa Turco, Cecilia |
| author_facet | Quintana, Maria Eugenia Barone, Lucas Forlenza, María Belén Trotta, Myrian Vanesa Turco, Cecilia Mansilla, Florencia Celeste Cardoso, Nancy Patricia Capozzo, Alejandra Victoria |
| author_sort | Quintana, Maria Eugenia |
| collection | INTA Digital |
| description | Low-cost high-throughput methods applicable to any virus strain are required for screening antiviral compounds against multiple field strains. Colorimetric cell-viability assays are used for this purpose as long as the viruses are cytopathic (CP) in cell culture. However, bovine viral diarrhoea virus (BVDV) strains circulating in the field are mostly non-cytopathic (NCP). An In Cell-ELISA aimed to measure viral infectivity by detecting a conserved protein produced during viral replication (non-structural protein 3, “NS3”) was developed. The ELISA is performed without harvesting the cells, directly on the 96-wells culture plate. NS3 In Cell-ELISA was tested for its ability to assess BVDV-specific antiviral activity of recombinant bovine type I and III IFNs. Results correlated to those measured by qRT-PCR and virus titration. NS3 In Cell-ELISA was also efficient in estimating the IC50 of two compounds with different antiviral activity. Estimation of the 50% inhibition dose of each IFN using six BVDV strains of different biotype and genotype showed that CP strains were more susceptible to both IFNs than NCP, while type 2 NCP viruses were more sensitive to IFN-I. The In Cell-ELISA format using a detector antibody against a conserved non-structural protein can be potentially applied to accurately measure infectivity of any
viral strain. |
| format | info:ar-repo/semantics/artículo |
| id | INTA6503 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA65032020-07-16T17:44:51Z A direct high-throughput in Cell-ELISA for measuring infectivity of cytopathic and non-cytopathic bovine viral diarrhoea virus strains applied to the assessment of antiviral activity Quintana, Maria Eugenia Barone, Lucas Forlenza, María Belén Trotta, Myrian Vanesa Turco, Cecilia Mansilla, Florencia Celeste Cardoso, Nancy Patricia Capozzo, Alejandra Victoria Bovine Viral Diarrhoea Pathogenicity Diarrea Viral Bovina Patogenicidad Cytopathic Cell-ELISA Low-cost high-throughput methods applicable to any virus strain are required for screening antiviral compounds against multiple field strains. Colorimetric cell-viability assays are used for this purpose as long as the viruses are cytopathic (CP) in cell culture. However, bovine viral diarrhoea virus (BVDV) strains circulating in the field are mostly non-cytopathic (NCP). An In Cell-ELISA aimed to measure viral infectivity by detecting a conserved protein produced during viral replication (non-structural protein 3, “NS3”) was developed. The ELISA is performed without harvesting the cells, directly on the 96-wells culture plate. NS3 In Cell-ELISA was tested for its ability to assess BVDV-specific antiviral activity of recombinant bovine type I and III IFNs. Results correlated to those measured by qRT-PCR and virus titration. NS3 In Cell-ELISA was also efficient in estimating the IC50 of two compounds with different antiviral activity. Estimation of the 50% inhibition dose of each IFN using six BVDV strains of different biotype and genotype showed that CP strains were more susceptible to both IFNs than NCP, while type 2 NCP viruses were more sensitive to IFN-I. The In Cell-ELISA format using a detector antibody against a conserved non-structural protein can be potentially applied to accurately measure infectivity of any viral strain. Instituto de Virología Fil: Quintana, María Eugenia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Tecnológicas; Argentina. Fil: Barone, Lucas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Fil: Forlenza, María Belén. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Fil: Trotta, Myrian Vanesa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Fil: Turco, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Fil: Mansilla, Florencia Celeste. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Fil: Cardoso, Nancy Patricia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Tecnológicas; Argentina. Fil: Capozzo, Alejandra Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Tecnológicas; Argentina. 2019-12-13T10:53:45Z 2019-12-13T10:53:45Z 2018-07-19 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion https://www.sciencedirect.com/science/article/pii/S0166093418301617 http://hdl.handle.net/20.500.12123/6503 0166-0934 https://doi.org/10.1016/j.jviromet.2018.07.010 eng info:eu-repo/semantics/restrictedAccess application/pdf Elsevier Journal of Virological Methods 260 (1): 75–81. (October 2018) |
| spellingShingle | Bovine Viral Diarrhoea Pathogenicity Diarrea Viral Bovina Patogenicidad Cytopathic Cell-ELISA Quintana, Maria Eugenia Barone, Lucas Forlenza, María Belén Trotta, Myrian Vanesa Turco, Cecilia Mansilla, Florencia Celeste Cardoso, Nancy Patricia Capozzo, Alejandra Victoria A direct high-throughput in Cell-ELISA for measuring infectivity of cytopathic and non-cytopathic bovine viral diarrhoea virus strains applied to the assessment of antiviral activity |
| title | A direct high-throughput in Cell-ELISA for measuring infectivity of cytopathic and non-cytopathic bovine viral diarrhoea virus strains applied to the assessment of antiviral activity |
| title_full | A direct high-throughput in Cell-ELISA for measuring infectivity of cytopathic and non-cytopathic bovine viral diarrhoea virus strains applied to the assessment of antiviral activity |
| title_fullStr | A direct high-throughput in Cell-ELISA for measuring infectivity of cytopathic and non-cytopathic bovine viral diarrhoea virus strains applied to the assessment of antiviral activity |
| title_full_unstemmed | A direct high-throughput in Cell-ELISA for measuring infectivity of cytopathic and non-cytopathic bovine viral diarrhoea virus strains applied to the assessment of antiviral activity |
| title_short | A direct high-throughput in Cell-ELISA for measuring infectivity of cytopathic and non-cytopathic bovine viral diarrhoea virus strains applied to the assessment of antiviral activity |
| title_sort | direct high throughput in cell elisa for measuring infectivity of cytopathic and non cytopathic bovine viral diarrhoea virus strains applied to the assessment of antiviral activity |
| topic | Bovine Viral Diarrhoea Pathogenicity Diarrea Viral Bovina Patogenicidad Cytopathic Cell-ELISA |
| url | https://www.sciencedirect.com/science/article/pii/S0166093418301617 http://hdl.handle.net/20.500.12123/6503 https://doi.org/10.1016/j.jviromet.2018.07.010 |
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