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Plaque assay for human coronavirus NL63 using human colon carcinoma cells
BACKGROUND: Coronaviruses cause a broad range of diseases in animals and humans. Human coronavirus (hCoV) NL63 is associated with up to 10% of common colds. Viral plaque assays enable the characterization of virus infectivity and allow for purifying virus stock solutions. They are essential for drug...
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2008
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2603006/ https://www.ncbi.nlm.nih.gov/pubmed/19014487 http://dx.doi.org/10.1186/1743-422X-5-138 |
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author | Herzog, Petra Drosten, Christian Müller, Marcel A |
author_facet | Herzog, Petra Drosten, Christian Müller, Marcel A |
author_sort | Herzog, Petra |
collection | PubMed |
description | BACKGROUND: Coronaviruses cause a broad range of diseases in animals and humans. Human coronavirus (hCoV) NL63 is associated with up to 10% of common colds. Viral plaque assays enable the characterization of virus infectivity and allow for purifying virus stock solutions. They are essential for drug screening. Hitherto used cell cultures for hCoV-NL63 show low levels of virus replication and weak and diffuse cytopathogenic effects. It has not yet been possible to establish practicable plaque assays for this important human pathogen. RESULTS: 12 different cell cultures were tested for susceptibility to hCoV-NL63 infection. Human colon carcinoma cells (CaCo-2) replicated virus more than 100 fold more efficiently than commonly used African green monkey kidney cells (LLC-MK2). CaCo-2 cells showed cytopathogenic effects 4 days post infection. Avicel, agarose and carboxymethyl-cellulose overlays proved suitable for plaque assays. Best results were achieved with Avicel, which produced large and clear plaques from the 4(th )day of infection. The utility of plaque assays with agrose overlay was demonstrated for purifying virus, thereby increasing viral infectivity by 1 log 10 PFU/mL. CONCLUSION: CaCo-2 cells support hCoV-NL63 better than LLC-MK2 cells and enable cytopathogenic plaque assays. Avicel overlay is favourable for plaque quantification, and agarose overlay is preferred for plaque purification. HCoV-NL63 virus stock of increased infectivity will be beneficial in antiviral screening, animal modelling of disease, and other experimental tasks. |
format | Text |
id | pubmed-2603006 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-26030062008-12-16 Plaque assay for human coronavirus NL63 using human colon carcinoma cells Herzog, Petra Drosten, Christian Müller, Marcel A Virol J Methodology BACKGROUND: Coronaviruses cause a broad range of diseases in animals and humans. Human coronavirus (hCoV) NL63 is associated with up to 10% of common colds. Viral plaque assays enable the characterization of virus infectivity and allow for purifying virus stock solutions. They are essential for drug screening. Hitherto used cell cultures for hCoV-NL63 show low levels of virus replication and weak and diffuse cytopathogenic effects. It has not yet been possible to establish practicable plaque assays for this important human pathogen. RESULTS: 12 different cell cultures were tested for susceptibility to hCoV-NL63 infection. Human colon carcinoma cells (CaCo-2) replicated virus more than 100 fold more efficiently than commonly used African green monkey kidney cells (LLC-MK2). CaCo-2 cells showed cytopathogenic effects 4 days post infection. Avicel, agarose and carboxymethyl-cellulose overlays proved suitable for plaque assays. Best results were achieved with Avicel, which produced large and clear plaques from the 4(th )day of infection. The utility of plaque assays with agrose overlay was demonstrated for purifying virus, thereby increasing viral infectivity by 1 log 10 PFU/mL. CONCLUSION: CaCo-2 cells support hCoV-NL63 better than LLC-MK2 cells and enable cytopathogenic plaque assays. Avicel overlay is favourable for plaque quantification, and agarose overlay is preferred for plaque purification. HCoV-NL63 virus stock of increased infectivity will be beneficial in antiviral screening, animal modelling of disease, and other experimental tasks. BioMed Central 2008-11-12 /pmc/articles/PMC2603006/ /pubmed/19014487 http://dx.doi.org/10.1186/1743-422X-5-138 Text en Copyright © 2008 Herzog et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Herzog, Petra Drosten, Christian Müller, Marcel A Plaque assay for human coronavirus NL63 using human colon carcinoma cells |
title | Plaque assay for human coronavirus NL63 using human colon carcinoma cells |
title_full | Plaque assay for human coronavirus NL63 using human colon carcinoma cells |
title_fullStr | Plaque assay for human coronavirus NL63 using human colon carcinoma cells |
title_full_unstemmed | Plaque assay for human coronavirus NL63 using human colon carcinoma cells |
title_short | Plaque assay for human coronavirus NL63 using human colon carcinoma cells |
title_sort | plaque assay for human coronavirus nl63 using human colon carcinoma cells |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2603006/ https://www.ncbi.nlm.nih.gov/pubmed/19014487 http://dx.doi.org/10.1186/1743-422X-5-138 |
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