Monitoring Influenza Virus Survival Outside the Host Using Real-Time Cell Analysis
| Autor: | India Leclercq, Quentin Grassin, Thomas Labadie, Christophe Batéjat, Jean-Claude Manuguerra |
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| Přispěvatelé: | Centre de Biologie Structurale [Montpellier] (CBS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Institut Pasteur [Paris] (IP), Cellule d'Intervention Biologique d'Urgence - Laboratory for Urgent Response to Biological Threats (CIBU), Environnement et Risques infectieux - Environment and Infectious Risks (ERI) |
| Rok vydání: | 2021 |
| Předmět: |
[SDV]Life Sciences [q-bio]
General Chemical Engineering Cell analysis Hemagglutinin Glycoproteins Influenza Virus Biology medicine.disease_cause Virus General Biochemistry Genetics and Molecular Biology Madin Darby Canine Kidney Cells Dogs Computer Systems Influenza A virus medicine Animals Infectious virus Infectivity Microbial Viability General Immunology and Microbiology Host (biology) Cell growth General Neuroscience Viral Load Cell biology Titer Kinetics Mutation Linear Models Virus Inactivation |
| Zdroj: | Journal of visualized experiments : JoVE Journal of visualized experiments : JoVE, 2021, 168, ⟨10.3791/61133⟩ |
| ISSN: | 1940-087X |
| Popis: | International audience; Methods for virus particle quantification represent a critical aspect of many virology studies. Although several reliable techniques exist, they are either time-consuming or unable to detect small variations. Presented here is a protocol for the precise quantification of viral titer by analyzing electrical impedance variations of infected cells in real-time. Cellular impedance is measured through gold microelectrode biosensors located under the cells in microplates, in which magnitude depends on the number of cells as well as their size and shape. This protocol allows real-time analysis of cell proliferation, viability, morphology and migration with enhanced sensitivity. Also provided is an example of a practical application by quantifying the decay of influenza A virus (IAV) submitted to various physicochemical parameters affecting viral infectivity over time (i.e., temperature, salinity, and pH). For such applications, the protocol reduces the workload needed while also generating precise quantification data of infectious virus particles. It allows the comparison of inactivation slopes among different IAV, which reflects their capacity to persist in given environment. This protocol is easy to perform, is highly reproducible, and can be applied to any virus producing cytopathic effects in cell culture. |
| Databáze: | OpenAIRE |
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