@article{b9ce97e9229a4d9a86c9cb85d004319a,
title = "An enzymatic virus-like particle assay for sensitive detection of virus entry",
abstract = "A viral entry assay where a beta-lactamase reporter protein fused to the influenza matrix protein-1 (BlaM1) is packaged as a structural component into influenza virus-like particles (VLPs) is described. The Bla reporter is released upon fusion with target cells and can be detected in live cells by flow cytometry, microscopy, or fluorometric plate reader for utility in high-throughput screening approaches. The production of BlaM1 VLPs and subsequent transfer of Bla activity to target cells requires the presence of influenza hemagglutinin (HA) and neuraminidase (NA). In addition, transfer of Bla by the VLPs can be blocked by an influenza neutralizing antibody, is impeded by a chemical inhibitor of influenza virus entry, and requires HA that is cleaved by a protease specific for its cleavage site. An analogous VLP system also was developed for Ebola (EBOV) and Marburg (MARV) viruses, demonstrating that this straightforward assay has broad application for studying the entry steps of enveloped viruses, especially those that require high levels of biosafety containment.",
keywords = "Ebola virus, Flow cytometry, Influenza virus, Marburg virus, Microscopy, Virus entry",
author = "Tscherne, {Donna M.} and Balaji Manicassamy and Adolfo Garc{\'i}a-Sastre",
note = "Funding Information: We thank Richard Cadagan and Osman Lizardo for technical support, Chris Basler for the 39E4 antibody, EBOV GP plasmid, and MARV GP plasmid, and Lijun Rong for the EBOV BlaVP40 plasmid. We also thank Osvaldo Martinez for useful discussions. Flow cytometry was performed at the Mount Sinai Flow Cytometry Shared Resource Facility. Microscopy was performed at the Mount Sinai Microscopy Shared Resource Facility, supported with funding from NIH-NCI shared resources grant 5R24 CA095823-04, NSF Major Research Instrumentation grant DBI-9724504 and NIH shared instrumentation grant 1 S10 RR0 9145-01. Research on influenza viruses in the Adolfo Garc{\'i}a-Sastre lab is partly funded by the Center for Research on Influenza Pathogenesis (CRIP), a National Institute for Allergy and Infectious Diseases (NIAID) sponsored Center of Excellence for Influenza Research and Surveillance HHSN266200700010C, by the Northeast Biodefense Center and by the Center for Investigating Viral Immunity and Antagonism (CIVIA) (NIAID grants U54AI57158 and U19AI62623, respectively), and by NIAID grants U01AI70469 and P01AI58113. Donna Tscherne is funded by NIAID grant 1F32AI081428.",
year = "2010",
month = feb,
doi = "10.1016/j.jviromet.2009.10.020",
language = "English",
volume = "163",
pages = "336--343",
journal = "Journal of Virological Methods",
issn = "0166-0934",
publisher = "Elsevier B.V.",
number = "2",
}