title: An enzymatic assay for detection of viral entry.
abstract: This unit describes a viral entry assay where a beta-lactamase reporter protein fused to the matrix protein of either influenza (BlaM1) or ebola virus (BlaVP40) is packaged as a structural component into virus-like particles (VLPs). The Bla reporter is released upon fusion with target cells and can be detected in live cells by flow cytometry, microscopy, or a fluorometric plate reader for utility in high-throughput screening approaches. The transfer of Bla to a target cell by BlaM1 or BlaVP40 VLPs requires the presence of influenza hemagglutinin (HA) and neuraminidase (NA) or EboV glycoprotein (GP), respectively. This straightforward assay has broad application for studying the entry steps of enveloped viruses, especially those that require high levels of biosafety containment.
authors.affiliation: Department of Microbiology, Mount Sinai School of Medicine, New York, New York, USA.
authors.forename: Donna M
authors.initials: DM
authors.lastname: Tscherne
authors.forename: Adolfo
authors.initials: A
authors.lastname: García-Sastre
mesh_terms: D002460:Cell Line; D029043:Ebolavirus / Q000502:physiology*; D057075:Enzyme Assays / Q000379:methods*; D017930:Genes, Reporter; D006801:Humans; D009975:Orthomyxoviridae / Q000502:physiology*; D014763:Viral Matrix Proteins / Q000235:genetics; D053586:Virus Internalization; D001618:beta-Lactamases / Q000235:genetics
chemical_list: C488936:M1 protein, Influenza A virus; D014763:Viral Matrix Proteins; D001618:beta-Lactamases