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Applied Veterinary Virology: The isolation and identification of viruses using cell cultures
Applied Veterinary Virology:
The isolation and identification of viruses using cell
cultures
Authors: Prof Estelle Venter
Licensed under a Creative Commons Attribution license.
FREQUENTLY ASKED QUESTIONS
1.
Can a cell culture laboratory be part of a microbiology laboratory?
Yes, but specific rooms separated from other diagnostic laboratories should be dedicated to
virology. If the laboratory receives samples from humans or samples containing highly
infectious agents, a virology unit with all the necessary safety precautions should be used.
2.
Is it possible to prepare your own cell culture medium or should one rather buy from a
reliable source?
It is always better to buy good quality medium but it is possible to produce your own medium
and do the sterility testing.
3.
Why is it necessary to quantify viruses?
For vaccine production, infection of animals in research projects and serology e.g. the serum
neutralization and haemagglutination inhibition tests, it is very important to know the
concentration of the virus.
4.
Can one use cell culture medium without using bovine serum?
Serum-free medium is commercially available but the effectivity of the medium for all types of
cells may vary. This is currently not commonly used by virology laboratories.
5.
Is serology always used to confirm the presence of a virus in a diagnostic sample?
The clinical signs and specific host species are a good indication of the cause of the disease.
Electron microscopy is also normally used, if available.
6.
Is trypsin the best enzyme to use during splitting of cells?
Yes – most virology laboratories are using trypsin.
7.
Which type of antimicrobial drug should one uses?
A broad spectrum, non-toxic antibiotic e.g. gentamycin. Penicillin and streptomycin are also
commonly used.
8.
How effective is the preparation and use of primary cell cultures?
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Applied Veterinary Virology: The isolation and identification of viruses using cell cultures
The preparation depends on the type and age of animal tissues used. Primary cultures are very
susceptible to isolate viruses especially from the same host species as the culture.
9.
Will all viruses give CPE?
Not all viruses produce CPE. Arboviruses do not produce CPE in insect cells and a mammalian
cell culture or another method should be used to confirm the presence of the virus. Noncytopathic bovine viral diarrhoea virus also e.g. will not produce CPE.
10.
Can all cell types grow in the same culture medium?
Most cells commonly used in a virology laboratory can grow in one basic medium but there are
cell types i.e. insect cells that need a specific formulation.
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