期刊
INFLUENZA AND OTHER RESPIRATORY VIRUSES
卷 9, 期 1, 页码 51-57出版社
WILEY
DOI: 10.1111/irv.12297
关键词
Airway epithelial cultures; influenzavirus B; respiratory viruses; VIDISCA-454; virus discovery
资金
- European Community [223498]
- VIDI grant from the Netherlands Organization for Scientific Research (NWO) [016.066.318]
- European Community's Sixth Framework Programme under the project GRACE, EC [LSHM-CT-2005-518226]
BackgroundCurrently, 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. MethodThree respiratory clinical samples were tested on well-differentiated pseudostratified tracheobronchial human airway epithelial (HAE) cultures grown at an air-liquid 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. ResultsInfected 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). ConclusionWe 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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据