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Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds
JoVE Journal
Biologia
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JoVE Journal Biologia
Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds

Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds

DOI:
10.3791/62903-v

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06:36 min

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September 08, 2021

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, , , , , , , ,
1CIRI, Centre International de Recherche en Infectiologie, GIMAP team,University of Lyon, INSERM U1111, CNRS, UMR5308, ENS Lyon, UCBL1, University of St-Etienne, France, 2Department of Infectious Agents and Hygiene,University Hospital of St-Etienne, St-Etienne, France, 3Department of Infectious Diseases,University Hospital of St-Etienne, St-Etienne, France, 4CIRI, Centre International de Recherche en Infectiologie, Staphylococcal Pathogenesis team,University of Lyon, INSERM U1111, CNRS UMR5308, ENS Lyon, UCBL1, University of Lyon, Lyon, France, 5Department of Bacteriology, Institute for Infectious Agents,Hospices Civils de Lyon, Lyon, France

Capítulos

  • 00:05Introduction
  • 00:41Determination of Cell Density and Viability
  • 01:33Preparation of the Bacterial Suspension
  • 02:32Cell Inoculation
  • 03:03Quantification of Intracellular Bacteria with Improved Enzyme Protection Assay (iEPA)
  • 04:44Results: Intracellular Staphylococcus aureus Load in Epithelial Cells
  • 05:54Conclusion

Summary

Tadução automática

Tadução automática

This protocol aims to describe how to study the extent of Staphylococcus aureus internalization and its ability to survive inside the human host cell, as well as the intracellular efficacy of antimicrobial compounds.

Tags

Keywords: Enzyme Protection AssayStaphylococcus AureusInternalizationIntracellular EfficacyAntimicrobial CompoundsA549 Epithelial CellsCell ViabilityBacterial SuspensionMOI (multiplicity Of Infection)Colony Counting

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