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Laboratory Virology Reading: Medical Microbiology; by Murray, Rosenthal, and Pfaller; 5th edition, (2005), chapter 51 I. Four main methods for diagnosis of viral infections A. Culture B. Serology 1. Test for immune response to virus C. Detection of viral antigens D. Detection of viral nucleic acids II. Cultivation A. Some viruses cannot be cultured B. Sampling: fluids, excreta, tissue, depending on symptoms and circumstances C. Laboratory animals D. Embryonated eggs E. Cell culture 1. Primary cell cultures 2. Diploid cell strains 3. Continuous cell lines 4. Recognition of viral growth in cell culture a. Cytopathic effect (CPE) b. Hemadsorbtion III. Cytology A. Cytopathic effect B. Inclusion bodies IV. Assay of viruses A. Biological 1. Plaque assay a. Principle b. Protocol 2. Transformation 3. Endpoint Method: TCID50, EID50, ID50, LD50 B. Physical, biochemical 1. Hemagglutination 2. Immunological tests for proteins 3. Assay for nucleic acid (Southern, PCR) 4. Enzymatic (reverse transcriptase for retroviruses) C. Comparison of quantitative methods V. Serology A. Neutralization, hemagglutination inhibition, ELISA, RIA, LA, western blot VI. Additional references A. Fields Virology, 4th edition, D.M. Knipe & P.M. Howley, eds., Lippincott Williams & Wilkins, 2001, Chapter 2

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