Title
Culturing of respiratory viruses in well-differentiated pseudostratified human airway epithelium as a tool to detect unknown virusesCulturing of respiratory viruses in well-differentiated pseudostratified human airway epithelium as a tool to detect unknown viruses
Author
Faculty/Department
Faculty of Medicine and Health Sciences
Research group
Vaccine & Infectious Disease Institute (VAXINFECTIO)
Publication type
article
Publication
Subject
Human medicine
Source (journal)
Influenza and other respiratory viruses
Volume/pages
9(2015):1, p. 51-57
ISSN
1750-2659
1750-2640
ISI
000346653800008
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
Background Currently, virus discovery is mainly based on molecular techniques. Here, we propose a method that relies on virus culturing combined with state-of-the-art sequencing techniques. The most natural ex vivo culture system was used to enable replication of respiratory viruses. Method Three respiratory clinical samples were tested on well-differentiated pseudostratified tracheobronchial human airway epithelial (HAE) cultures grown at an airliquid interface, which resemble the airway epithelium. Cells were stained with convalescent serum of the patients to identify infected cells and apical washes were analyzed by VIDISCA-454, a next-generation sequencing virus discovery technique. Results Infected cells were observed for all three samples. Sequencing subsequently indicated that the cells were infected by either human coronavirus OC43, influenzavirus B, or influenzavirus A. The sequence reads covered a large part of the genome (52%, 82%, and 57%, respectively). Conclusion We present here a new method for virus discovery that requires a virus culture on primary cells and an antibody detection. The virus in the harvest can be used to characterize the viral genome sequence and cell tropism, but also provides progeny virus to initiate experiments to fulfill the Koch's postulates.
Full text (open access)
https://repository.uantwerpen.be/docman/irua/5423c6/9ac063ec.pdf
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