Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model

Bridget S. Banach, Jan M. Orenstein, Linda M. Fox, Scott H. Randell, Anne H. Rowley, Susan C. Baker*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)19-26
Number of pages8
JournalJournal of Virological Methods
Volume156
Issue number1-2
DOIs
StatePublished - Mar 2009

Keywords

  • Coronavirus
  • HAE culture
  • HCoV-NL63
  • TEM
  • Whole transcriptome amplification

ASJC Scopus subject areas

  • Virology

Fingerprint Dive into the research topics of 'Human airway epithelial cell culture to identify new respiratory viruses: Coronavirus NL63 as a model'. Together they form a unique fingerprint.

Cite this