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laboratory virology

Diagnosis of viral infections. CultureDetection of viral nucleic acidsDetection of viral antigensSerologyTest for immune response to virus. Virus Culture. Some viruses cannot be culturedSamplingfluids, excreta, tissue, depending on symptoms and circumstancesLaboratory animalsEmbryonated eggsCell culturePrimary cell culturesDiploid cell strainsContinuous cell linesTransformation.

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laboratory virology

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    1. Laboratory virology

    3. 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

    6. Primary cell culture

    7. Subculture

    9. 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

    10. Transformation Immortalization Loss of contact inhibition Anchorage independence Growth in soft agar Growth in suspension Tumor formation in athymic (nude) mice

    11. Recognition of viral growth in cell culture Cytopathic effect (CPE) Morphological changes Inclusion bodies Hemadsorbtion

    13. CPE: vaccinia

    15. CPE: inclusion bodies

    16. Hemadsorbtion

    18. Assay of viruses Biological Plaque assay Transformation Endpoint Method: TCID50, EID50, ID50, LD50 Physical, biochemical Hemagglutination Direct particle count Immunological tests for proteins Assay for nucleic acid (Southern, PCR) Enzymatic (reverse transcriptase for retroviruses) Comparison of quantitative methods

    19. Plaque formation

    20. Plaque

    21. Plaque formation

    23. Plaque assay

    24. Transformationloss of contact inhibition

    27. Hemagglutination

    30. Direct particle count

    34. Serology Neutralization Hemagglutination inhibition Western blot ELISA, radioimmune assay (RIA)

    38. Summary Four main clinical diagnostic techniques Culture, serology, antigen detection, nucleic acid detection Virus culture Not all viruses can be cultured Cultured cell types Cytopathic effect Virus quantitation Biological Physical Basic serological techniques

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