9-14 1PM Laboratory virology

9-14 1PM Laboratory virology - Laboratory virology...

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Laboratory virology
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Diagnosis of viral infections Culture Detection of viral nucleic acids Detection of viral antigens Serology Test for immune response to virus
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Virus Culture Some viruses cannot be cultured Sampling fluids, excreta, tissue, depending on symptoms and circumstances Laboratory animals Embryonated eggs Cell culture Primary cell cultures Diploid cell strains Continuous cell lines Transformation
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From Medical Microbiology, 5 th ed., Murray, Rosenthal & Pfaller, Mosby Inc., 2005, Table 51-1. Specimens for viral diagnosis
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Growth of virus on embryonated eggs Davis, Duylbecco, Eisen, Ginsberg “Microbiology” 4 th ed, J.B. Lippincott 1990, Fig. 48-1
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Primary cell culture + enzymes time
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Subculture enzymes time
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Growth of cells in culture. A primary culture is defined as the original plating of cells from a tissue, grown to a confluent monolayer, without subculturing. A cell strain (solid line) is defined as a euploid population of cells subcultivated more than once in vitro, lacking the property of indefinite serial passage. Cell strains ultimately undergo degeneration and death, also called crisis or senescence. A cell line (dashed line) is an aneuploid population of cells that can be grown in culture indefinitely. Spontaneous transformation or alteration of a cell strain to an immortal cell line can occur at any time during cultivation of the cell strain. The time in culture and corresponding number of subcultivations or passages are shown on the abscissas. The ordinate shows the total number of cells that would accumulate if all were retained in culture. (From Fields Vriology (2007) 5th edition, Knipe, DM & Howley, PM, eds, Wolters Kluwer/Lippincott Williams & Wilkins, Philadelphia Fig. 2.3) Cell culture
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Cultured cells Primary Heterogeneous – many cell types Closest to animal Technical hassle Diploid cell strain Relatively homogeneous – fewer cell types Further from animal Technically less hassle Continuous cell line Immortal Most homogeneous Genetically weird – furthest from animal Hassle free Suspension or monolayer
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Transformation Immortalization Loss of contact inhibition Anchorage independence Growth in soft agar Growth in suspension Tumor formation in athymic (nude) mice
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Recognition of viral growth in cell culture Cytopathic effect (CPE) Morphological changes Inclusion bodies Hemadsorbtion
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