Isolation and identification of viruses using cell cultures

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Schematic presentation of the haemagglutination test

Schematic presentation of the haemagglutination test

Term: 2014
Published: February 26, 2014
Revised: March 28, 2014

This submodule broadly covers the use of cell cultures for the isolation and identification of animal viruses and the use of different media for the propagation of different cell types.

You will learn about laboratory equipment and basic and additional requirements to set up a veterinary diagnostic virology laboratory. The types of biological safety cabinets and their functions are addressed and the requirement for absolute sterility emphasized as a key feature. The nomenclature of cell lines and types of cells and the different media to sustain growth are described. Cell culture media and the use of serum and antimicrobial drugs are included.

You will be taken through techniques for establishing primary cultures and continuous cell lines, the use of enzymes to dissociate tissues and cell line sub-culturing and storage or cryopreservation of cells. Different cell lines and the viruses they support are discussed. Diagnostic sample inoculation and maintenance and harvesting of cultures are described. Short, medium and long term storage of viruses using storage medium and antibiotics is covered.

Confirmation and demonstration of viruses from the preparation of specimens to identification techniques is covered. Cytopathic effects caused by viruses are emphasized and laboratory techniques such as serum virus neutralization, haemadsorption, haemagglutination and haemagglutination inhibition are explained.
Quantification of viruses using the plaque forming assay method as well as a dose response curve produced in either cell cultures, laboratory animals or embryonated chicken eggs and calculation of the 50% endpoint, is described.

 

About The Instructor

Prof Estelle Venter

Prof Estelle Venter

  • BSc, MSc, PhD
  • Professor: Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, South Africa
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This Work, Isolation and identification of viruses using cell cultures, by Prof Estelle Venter is licensed under a Creative Commons Attribution license.