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.

我们最近确定人类诺如病毒 (HuNoV) 在将临床样本接种到蛋黄后在斑马鱼幼虫中有效复制，从而提供了一个简单而强大的体内系统来研究 HuNoV。在此协议扩展中，我们详细描述了通过显微注射接种病毒、随后的日常监测和幼虫收获，然后是病毒 RNA 量化。该协议可用于研究基因组 (G)I 和 GII HuNoV 在体内 3-4 d 内的病毒复制。此外，我们描述了如何在多孔板中使用受 HuNoV 感染的斑马鱼幼虫评估小分子的体内抗病毒作用和毒性，而无需特定的配方。这对药物发现工作构成了巨大的优势，