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Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model
Propagation of new human respiratory virus pathogens in established cell lines is hampered by a lack of predictability regarding cell line permissivity and by availability of suitable antibody reagents to detect infection in cell lines that do not exhibit significant cytopathic effect. Recently, mol...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Elsevier B.V.
2009
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2671689/ https://www.ncbi.nlm.nih.gov/pubmed/19027037 http://dx.doi.org/10.1016/j.jviromet.2008.10.022 |
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author | S. Banach, Bridget Orenstein, Jan M. Fox, Linda M. Randell, Scott H. Rowley, Anne H. Baker, Susan C. |
author_facet | S. Banach, Bridget Orenstein, Jan M. Fox, Linda M. Randell, Scott H. Rowley, Anne H. Baker, Susan C. |
author_sort | S. Banach, Bridget |
collection | PubMed |
description | Propagation of new human respiratory virus pathogens in established cell lines is hampered by a lack of predictability regarding cell line permissivity and by availability of suitable antibody reagents to detect infection in cell lines that do not exhibit significant cytopathic effect. Recently, molecular methods have been used to amplify and identify novel nucleic acid sequences directly from clinical samples, but these methods may be hampered by the quantity of virus present in respiratory secretions at different time points following the onset of infection. Human airway epithelial (HAE) cultures, which effectively mimic the human bronchial environment, allow for cultivation of a wide variety of human respiratory viral pathogens. The goal of the experiments described here was to determine if propagation and identification of a human respiratory virus may be achieved through inoculation of HAE cultures followed by whole transcriptome amplification (WTA) and sequence analysis. To establish proof-of-principle human coronavirus NL63 (HCoV-NL63) was evaluated, and the first visualization of HCoV-NL63 virus by transmission electron microscopy (TEM) is reported. Initial propagation of human respiratory secretions onto HAE cultures followed by TEM and WTA of culture supernatant may be a useful approach for visualization and detection of new human respiratory pathogens that have eluded identification by traditional approaches. |
format | Text |
id | pubmed-2671689 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-26716892010-03-01 Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model S. Banach, Bridget Orenstein, Jan M. Fox, Linda M. Randell, Scott H. Rowley, Anne H. Baker, Susan C. J Virol Methods Article Propagation of new human respiratory virus pathogens in established cell lines is hampered by a lack of predictability regarding cell line permissivity and by availability of suitable antibody reagents to detect infection in cell lines that do not exhibit significant cytopathic effect. Recently, molecular methods have been used to amplify and identify novel nucleic acid sequences directly from clinical samples, but these methods may be hampered by the quantity of virus present in respiratory secretions at different time points following the onset of infection. Human airway epithelial (HAE) cultures, which effectively mimic the human bronchial environment, allow for cultivation of a wide variety of human respiratory viral pathogens. The goal of the experiments described here was to determine if propagation and identification of a human respiratory virus may be achieved through inoculation of HAE cultures followed by whole transcriptome amplification (WTA) and sequence analysis. To establish proof-of-principle human coronavirus NL63 (HCoV-NL63) was evaluated, and the first visualization of HCoV-NL63 virus by transmission electron microscopy (TEM) is reported. Initial propagation of human respiratory secretions onto HAE cultures followed by TEM and WTA of culture supernatant may be a useful approach for visualization and detection of new human respiratory pathogens that have eluded identification by traditional approaches. Elsevier B.V. 2009-03 2008-12-05 /pmc/articles/PMC2671689/ /pubmed/19027037 http://dx.doi.org/10.1016/j.jviromet.2008.10.022 Text en Copyright © 2008 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article S. Banach, Bridget Orenstein, Jan M. Fox, Linda M. Randell, Scott H. Rowley, Anne H. Baker, Susan C. Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model |
title | Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model |
title_full | Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model |
title_fullStr | Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model |
title_full_unstemmed | Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model |
title_short | Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model |
title_sort | human airway epithelial cell culture to identify new respiratory viruses: coronavirus nl63 as a model |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2671689/ https://www.ncbi.nlm.nih.gov/pubmed/19027037 http://dx.doi.org/10.1016/j.jviromet.2008.10.022 |
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