Abstract
We have recently established that human norovirus (HuNoV) replicates efficiently in zebrafish larvae after inoculation of a clinical sample into the yolk, providing a simple and robust in vivo system in which to study HuNoV. In this Protocol Extension, we present a detailed description of virus inoculation by microinjection, subsequent daily monitoring and harvesting of larvae, followed by viral RNA quantification. This protocol can be used to study viral replication of genogroup (G)I and GII HuNoVs in vivo within 3–4 d. Additionally, we describe how to evaluate the in vivo antiviral effect and toxicity of small molecules using HuNoV-infected zebrafish larvae, in multi-well plates and without the need for specific formulations. This constitutes a great advantage for drug discovery efforts, as no specific antivirals or vaccines currently exist to treat or prevent norovirus gastroenteritis.
| Original language | English |
|---|---|
| Journal | Nature Protocols |
| Volume | 16 |
| Issue number | 4 |
| Pages (from-to) | 1830-1849 |
| Number of pages | 20 |
| ISSN | 1754-2189 |
| DOIs | |
| Publication status | Published - 04.2021 |
Funding
We very much appreciate the expert technical assistance and dedication of J. Rymenants, L. Bervoets and J. Maes. We thank the KU Leuven aquatic facility for breeding the zebrafish larvae. We thank the pediatrics department and the CEMOL Molecular Diagnostic department of the University Hospital of Leuven for the collaboration. J.V.D. and the research leading to these results has received funding from the Scientific Fund for Research of Flanders (FWO) as an SB doctoral fellow. A.C. is supported by internal funding from the KU Leuven.
Research Areas and Centers
- Centers: Center for Structural and Cell Biology (CSCM/ZMSZ)
DFG Research Classification Scheme
- 2.21-04 Virology
