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Virology
Definition of Virus
Size
Methods of analysis
2. Ultracentrifuge
3. Electron microscope
4. X-ray crystallography
Size of Viruses
Virion
Capsid
The protein shell, or coat, that encloses the nucleic acid genome.
Functions:
a. Protect the viral nucleic acid.
b. Participate in the viral infection.
c. Antigenic and specific for each virus type
d. Provides structural symmetry to the virus particle
Nucleocapsid
Capsid
Viral core
DNA RNA
Single Stranded + or -
1. Cubical/Icosahedral
(Adeno virus, Coxsackie virus, CMV, EBV, Hepatitis virus, HSV,
Polio virus, Rubella virus)
2. Helical
(Influenza, Rubeola, Mumps, Rabies, Hanta, Corona viruses)
3. Complex
(Bacteriophage, Pox viruses)
Cubic or icosahedral symmetry
Complex symmetry
Naked
Virus
Enveloped
Virus
Shape of viruses
The viral multiplication cycle can be divided into six sequential phases,
though the phases may sometimes be overlapping
1. Adsorption or attachment
2. Penetration
3. Uncoating
4. Biosynthesis
5. Maturation
6. Release
Adsorption
Virions may come into contact with cells by random collision but
adsorption takes place only if there is an affinity between the two
Penetration
Virus particles may be engulfed by a mechanism resembling
phagocytosis, a process known as viropexis
In the case of enveloped viruses, the viral envelope may fuse with
the plasma membrane of the host cell and release the nucleocapsid
into the cytoplasm
Uncoating
Process of stripping the the virus of its outer layers and capsid
so that the nucleic acid is released into the cell
Biosynthesis
During this phase, viral nucleic acid, capsid protein, enzymes
necessary in the various stages of viral synthesis, assembly
and release will be synthesised
Virion assembly may take place in the host cell nucleus (Herpes and
adenoviruses) or cytoplasm (picorna and poxviruses)
Release
Progeny of bacterial viruses release by the lysis of the infected bacterium.
In the case of animal viruses, release usually occurs without cell lysis
Incomplete viruses
Incomplete viruses are seen in large proportions, when cells are
infected with a high dose (MOI) of the influenza virus
The virus yield will have a high hemagglutinin titer but low infectivity
and this is known as the von Magnus phenomenon
Abortive infection
Some viruses are genetically defective in that when they infect cells,
they are unable to give rise to fully formed progeny
Human volunteers
Animal inoculation
Embryonated eggs
Tissue culture
a) Organ culture
b) Explant culture
c) Cell culture
Animal inoculation
The embryonated egg offers several sites for the cultivation of viruses
Inoculation into the amniotic sac is employed for the primary isolation
of the influenza virus
Organ culture
Small bits of organs can be maintained in vitro for days and weeks,
preserving their original architecture and function
Explant culture
Although embryonated eggs and laboratory animals are very useful for
isolation viruses, cell culture is the sole system for virus isolation in
most laboratories
Useful for the isolation of viruses and their cultivation for vaccine
production
These are cells of a single type, usually derived from cancer cells
and capable of continuous serial cultivation indefinitely
(eg: HeLa, HEp-2, Vero, KB cell lines)
Cytopathic effect
Many viruses cause morphological changes in cultured cells, these
changes can be readily observed by microscopic examination and
these changes are known as cytopathic effects (CPE)
Hemadsorption
When hemagglutinating viruses grow in cell culture, their presence
can be indicated by the addition of guinea pig erythrocytes to the
cultures
Interference
The growth of a non-cytopathogenic virus in cell culture can be tested
by the subsequent challenge with a known cytopathogenic virus
Transformation
Oncogenic viruses induce cell transformation and loss of contact
inhibition, so that growth appears in a piled-up fashion producing
microtumors
Immunofluorescence
Viral assay
The virus content of a specimen can be assayed in two ways
a)Quantal assays
Only indicate the presence or absence of infectious viruses but using
serial dilutions of virus suspensions and with the aid of statistical
methods, reasonably accurate estimates of infectivity can be obtained
End points used for infectivity titration are the death of the animal,
production of hemagglutinin in allantoic fluid or the appearance of
CPE in cell cultures
1.Plaque assay
A viral suspension is added to a monolayer of cultured cells in a bottle
or petri dish, and after adsorption, the medium is removed and
replaced with a solid agar gel
2. Pock assay
Plaque assay Pock assay
Classification and nomenclature of viruses
Viruses are broadly classified into DNA and RNA viruses and then
further divided into families, subfamilies, genera and species
Viroids
Prions