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By,
A. Raihanathus Sahdhiyya
I M.Sc. Microbiology
 History
Techniques in virology
Virus cultivation method
 late 19th century  the independent work of Dimitri
Ivanofsky and Martinus Beijerinck marked the beginning of the
field of virology.
Causative agent in tobacco plant disease  able to pass
through the filter retaining Bacteria
In 1898,
Friedrich Loeffler and Paul Frosch applied the filtration criteria
to a disease in cattle known as foot and mouth disease.
1. Detection of a meaningful immune response by immunologic
assay
2. Identification of the agent by staining (light and electron
microscopy)
3. Isolation and identification of the agent (cell culture or fertile
eggs)
4. Detection of viral nucleic acid (probes or amplification).
• Virus neutralization (VN)
• Hemagglutination inhibition (HI)
• Enzyme linked immunosorbent assay (ELISA)
• Indirect fluorescent antibody (IFA)
• Complement fixation (CF)
• Agar-gel immunodiffusion (AGID)
• Agar-gel precipitin (AGP)
• Latex agglutination (LA).
sample is incubated with the target virus
changes in cell culture are observed
(cytopathic effect)
contains antibodies no antibodies
Prevent viral growth viral growth
no CPE CPE
Light Microscopy
Viruses - too small to be seen using a standard light microscope
Antibodies labeled with an indicator
visualization of the virus cluster
peroxidase
light microscope ultraviolet (UV) light microscope
fluorescence
• The magnification -50,000x magnified  provides the ability to see the
viral particles
• Problem :
lack of sensitivity (1,000,000 virus particles/ ml of fluid is
required )
To improve this, immune electron microscopy
sample is incubated with an antibody
antibody-antigen reaction results in clumps
of the virus
easier visualization.
Virology techniques
Virology techniques
Virology techniques
Virology techniques
Virology techniques
Virology techniques

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Virology techniques

  • 1. By, A. Raihanathus Sahdhiyya I M.Sc. Microbiology  History Techniques in virology Virus cultivation method
  • 2.  late 19th century  the independent work of Dimitri Ivanofsky and Martinus Beijerinck marked the beginning of the field of virology. Causative agent in tobacco plant disease  able to pass through the filter retaining Bacteria In 1898, Friedrich Loeffler and Paul Frosch applied the filtration criteria to a disease in cattle known as foot and mouth disease.
  • 3. 1. Detection of a meaningful immune response by immunologic assay 2. Identification of the agent by staining (light and electron microscopy) 3. Isolation and identification of the agent (cell culture or fertile eggs) 4. Detection of viral nucleic acid (probes or amplification).
  • 4. • Virus neutralization (VN) • Hemagglutination inhibition (HI) • Enzyme linked immunosorbent assay (ELISA) • Indirect fluorescent antibody (IFA) • Complement fixation (CF) • Agar-gel immunodiffusion (AGID) • Agar-gel precipitin (AGP) • Latex agglutination (LA).
  • 5. sample is incubated with the target virus changes in cell culture are observed (cytopathic effect) contains antibodies no antibodies Prevent viral growth viral growth no CPE CPE
  • 6.
  • 7.
  • 8. Light Microscopy Viruses - too small to be seen using a standard light microscope Antibodies labeled with an indicator visualization of the virus cluster peroxidase light microscope ultraviolet (UV) light microscope fluorescence
  • 9. • The magnification -50,000x magnified  provides the ability to see the viral particles • Problem : lack of sensitivity (1,000,000 virus particles/ ml of fluid is required ) To improve this, immune electron microscopy sample is incubated with an antibody antibody-antigen reaction results in clumps of the virus easier visualization.