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Cultivation of Viruses

Cultivation of Viruses. By. Dr. Emad AbdElhameed Morad. Lecturer of Medical Microbiology and Immunology. Viruses are obligate intracellular organisms. So, they must be cultivated on living cells. So, viruses could be cultivated on either:. Cell culture. Embryonated eggs.

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Cultivation of Viruses

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  1. Cultivation of Viruses By Dr. Emad AbdElhameed Morad Lecturer of Medical Microbiology and Immunology

  2. Viruses are obligate intracellular organisms. • So, they must be cultivated on living cells. • So, viruses could be cultivated on either: Cell culture Embryonated eggs Laboratory animals

  3. Cell culture • Cell culture monolayer is the most commonly used system for isolation of viruses. • Cell culture has three types: • Primary cell line: • Prepared for example from monkey kidney. • These cells can divide for 4-6 passages then die. • Human diploid cell line: • Prepared from human embryo fibroblasts. • These cells can divide for 50 passages then die. • Continuous cell line: • Prepared from tumor cells. • Example, HeLa cell line derived from carcinoma of the cervix. • These cells can divide indefinitely.

  4. Detection of viral growth in tissue culture • Cytopathic effect • Plaque formation • Inclusion bodies • Hemadsorption • Interference

  5. Cytopathic effect (CPE) • They are changes which occur in the cells of the cell culture. • Examples: • Cell death. • Rounding and grape like cluster formation. • Syncytium formation (multinucleated giant cell formation). • Cell transformation (the cells become malignant cells).

  6. Plaque formation • Plaques are virally infected areas in the cell culture. • Each plaque is produced by a single virus. • These plaques are seen by the naked eyeas unstained areas when using vital stains as neutral red.

  7. Inclusion bodies • They are sites of assembly of the virus components inside the infected cells. • May be intranuclear or intracytoplasmic. • Could be seen by the light microscope.

  8. Hemadsorption • When red blood cells are added to the infected cells, they will appear as rosettes. • This is useful in case of influenza and mumps viruses.

  9. Interference • Some viruses do not produce CPE on cell culture. • However, the growth of these viruses on cell culture is detected by interference with the formation of CPE by other viruses.

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