Karlsson Erik A, Meliopoulos Victoria A, Tran Vy, Savage Chandra, Livingston Brandi, Schultz-Cherry Stacey, Mehle Andrew
Department of Infectious Diseases, St Jude Children's Research Hospital, Memphis, TN, USA.
Virology Unit, Institut Pasteur du Cambodge, Phnom Penh, Cambodia.
Methods Mol Biol. 2018;1836:431-459. doi: 10.1007/978-1-4939-8678-1_21.
To streamline standard virological assays, we developed bioluminescent replication-competent influenza reporter viruses that mimic their parental counterparts. These reporter viruses provide a rapid and quantitative readout of viral infection and replication. Moreover, they permit real-time in vivo measures of viral load, tissue distribution, and transmission in the same cohort of animals over the entire course of infection-measurements that were not previously possible. Here we provide detailed protocols using bioluminescent reporter viruses for in vivo imaging in mice and ferrets. We also describe cell culture-based techniques using reporter viruses for quantification of viral titers and performing microneutralization assays. The ease, speed, and adaptability of these approaches have the potential to accelerate multiple areas of influenza virus research.
为了简化标准病毒学检测方法，我们开发了具有生物发光能力且能复制的流感报告病毒，这些病毒可模拟其亲本病毒。这些报告病毒能对病毒感染和复制进行快速定量检测。此外，它们还能在感染全过程中，对同一批动物体内的病毒载量、组织分布及传播情况进行实时检测，而这些检测此前是无法实现的。在此，我们提供了使用生物发光报告病毒对小鼠和雪貂进行体内成像的详细方案。我们还描述了基于细胞培养的技术，利用报告病毒定量病毒滴度并进行微量中和试验。这些方法的简便性、快速性和适应性有潜力加速流感病毒研究的多个领域。
