Unit

UNIT 26.12 An Enzymatic Assay for Detection of Viral Entry

  1. Donna M. Tscherne1,
  2. Adolfo García-Sastre1,2,3

Published Online: 1 JUN 2011

DOI: 10.1002/0471143030.cb2612s51

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Tscherne, D. M. and García-Sastre, A. 2011. An Enzymatic Assay for Detection of Viral Entry. Current Protocols in Cell Biology. 51:26.12:26.12.1–26.12.10.

Author Information

  1. 1

    Department of Microbiology, Mount Sinai School of Medicine, New York, New York

  2. 2

    Department of Medicine, Division of Infectious Diseases, Mount Sinai School of Medicine, New York, New York

  3. 3

    Global Health and Emerging Pathogens Institute, Mount Sinai School of Medicine, New York, New York

Publication History

  1. Published Online: 1 JUN 2011
  2. Published Print: JUN 2011

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. Curr. Protoc. Cell Biol. 51:26.12.1-26.12.10. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • virus entry;
  • influenza virus;
  • ebola virus;
  • Marburg virus;
  • flow cytometry;
  • microscopy