General virology

A Virus is an obligate intracellular infectious agent usually beyond the resolution of light
microscope and is minimally constructed of a genome consisting of either RNA or DNA
and a protective protein coat (capsid). Many viruses have additional structures
(envelope, matrix protein, glycoprotein spikes etc).
Virion is a complete virus particle; it may be enveloped or non-enveloped (naked).
Size ranges between 20-300nm.
Shapes may resemble brick, rod, sphere, etc.
Structure of a virion:

1. Genome: virus contains either RNA or DNA as their nucleic acid, but never both.
Attached to nucleic acid are some nucleoproteins (enzymes). Viral genome is
haploid, except retrovirus that has diploid nucleic acid

• DNA is always double stranded except that of parvovirus

• RNA is always single stranded except that of reovirus
• DNA is always entire
• RNA may be entire or segmented (influenza virus)

2. Capsid is a protein coat that surrounds the nucleic acid. It is made up of subunits
called capsomers. Capsid surrounds the nucleic acid core symmetrically. The
capsid symmetry is of following two types:
a) Icosahedral - Here capsomers are arranged in 20 triangles and give the virion a
spherical outline.
b) Helical - Here capsomers are arranged in a hollow coil giving a rod-shaped
outline to the virus.

Functions of capsid:
• Protect nucleic acid core
• Attach to host cell surface (non-enveloped virus).
• Stimulate production of neutralizing antibody and activate virus specific Tc cell
(nonenveloped virus).

1. Matrix and tegument proteins- these proteins fill the space between capsid and
envelope.
Functions:
• Matrix proteins mediate interaction between capsid and envelope.
• Tegument proteins are regulatory proteins.

2. Envelope is a lipoprotein membrane, that lie outside capsid. Lipid is derived from
host membrane during budding and protein is virus specific. Presence of envelope
makes a virus more fragile. Enveloped viruses are vulnerable to drying, detergents,
extremes of pH and temperature. Projecting from the envelope are glycoprotein
spikes.

Functions of glycoprotein spikes:

• help in attachment to host cell.
• Neutralizing antibody is produced against it.

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Viruses are obligate intracellular parasite: They lack protein and energy generating
system and depend on host cell machineries for producing protein and energy.
(Obligate intracellular bacteria are chlamydia and rickettsia)

Virus is not a cell as it does not have nucleus, cytoplasm, and cytoplasmic membrane.
They are beyond resolution of light microscope.

Properties of virus:
It has either DNA or RNA but never both.
It does not have any organelle.
It does not reproduce by binary fission or mitosis.
It cannot be seen by light microscope (seen by electron microscope).
It is filtrable

Viruses differ from bacteria in that they are:

1) Smaller and cannot be seen by light microscope (Pox virus can be seen by light
microscope as featureless particle).
2) Possess either DNA or RNA but never both
3) Cannot synthesize its protein and energy (as virus is devoid of organelles).
4) Multiply by replication not binary fission.
5) Do not grow in cell free or inanimate media (grow in tissue culture media,
embryonated egg etc.).
6) Resistant to antibiotic
7) Sensitive to interferon

Replication: Virus reproduces by replication. Unlike cells, where two daughter cells are
produced from parent cell, one virus produces many viruses at a time.
At first virus disassemble, produces many copies of nucleic acid and proteins, then
assemble to produce an entire virion, and released from infected cell.

DNA virus replicates in nucleus (except pox virus)

RNA virus replicates in cytoplasm (except – Retro, HDV, and influenza virus)

Steps of replication:
1. Adsorption by capsid or glycoprotein spikes to a specific host cell receptor.
2. Penetration into host cell by:
• Receptor mediated endocytosis
• Fusion with host cell membrane
• Direct penetration (poliovirus, bacteriophage)
3. Uncoating, means release of viral nucleic acid from capsid
4. Early transcription - synthesis of viral mRNA
5. Early translation - synthesis of enzymatic proteins
6. Genome replication
7. Late transcription
8. Late translation - synthesis of structural proteins
9. Assembly
10. Release from host cell by:
• Lysis
• Budding. During budding a virus acquires envelope from
host cell membrane or nuclear membrane.

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Classification of virus:
Virus may be classified based on
1. Nucleic acid
• DNA virus: Adenovirus, Pox virus, Herpes virus etc.
• RNA virus: Retrovirus, Corona virus, Poliovirus etc.
2. Envelope
• Enveloped virus: Corona virus, Retrovirus, Herpes virus etc. Presence of envelope
makes a virus more vulnerable to drying, detergents, extreme pH.
• Naked or nonenveloped virus: Adenovirus, Poliovirus etc.

There is a correlation between presence of virus and transmission. It has been

observed that virtually all virus that are transmitted by fecal oral route are
nonenveloped. On the other hand, most, enveloped virus is transmitted by close contact
like blood or sexual contact.

Defective virus is one that lacks one or more functional genes required for viral
replication. They require helper activity from another virus for replication.
Types:
Defective interfering particle: during the growth of most human viruses, many more
defective than infectious particles are produced. They may interfere with the growth of
the infectious particles.
• HDV is another type of defective virus and requires help from unrelated HBV for
replication.
• Pseudovirion: are defective virus particle that contain host cell DNA rather than
viral genome. They can infect cell but cannot replicate.

Atypical virus-like agents

1. Viroids are infectious agents, solely composed of single molecule of circular RNA
without protein coat or envelope. They cause several plant diseases.

2. Prions are infectious particle composed solely of protein coded by cellular gene.
They cause disease in both human and animal. The diseases are characterized by:
• Long incubation period
• Gradual onset
• Progressive but invariably fatal course

Examples of some prion-mediated diseases are - Kuru, Creutzfeld-Jacob disease.

Comparison between Prion and Virion

Features Prion Conventional viruses

Nucleic acid absent present
Protein Present encoded by cellular Present encoded by viral
genes genes
Appearance under electron Filamentous rod. (Amyloid Icosahedral or helical
microscope like) symmetry.
Infection induces antibody no yes
Inflammation no yes
Inactivated rapidly by UV No yes

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Bacriophage is a virus that replicates inside bacteria. Bacteriophage infected bacteria
may have two effects:
a) Lysis of infected bacteria
b) Lysogenic conversion. It refers to when the infected bacteria acquire a new property
such as production of exotoxin. These exotoxins are called phage-mediated
exotoxins.
Following are some examples of phage-mediated exotoxins:
a. Cholera toxin
b. Erythrogenic toxin
c. Botulinum toxim
d. Diphtheria toxin
e. Verotoxin
f. Shiga toxin

Pathogenesis of viral disease:

1. Destruction of cell with loss of its function.
Infected cell may be destroyed by:
• direct effect of virus (Herpes virus)
• by cytotoxic T cell (Hepatitis virus)

2. Growth retardation- some virus reduces the rate of cell duplication. It has no effect in
adult. In rapidly growing tissue (fetus) reduction in rate of cell multiplication result
in incomplete tissue or organ development (rubella virus). It has no effect in adult.

3. Oncogenic effect - some virus causes malignant transformation.

Oncogenic viruses/cancer producing virus.

RNA
a. HCV-hepatocellular carcinoma.
b. Human T cell lymphotrophic virus- leukaemia, lymphoma.
DNA
a. Human papilloma virus-carcinoma cervix (vaccine is available).
b. EBV-Burkitts lymphoma.
- Nasopharyngeal ca.
- Thymic ca.
- B cell lymphoma.
c. Human herpes 8 - Kaposis sarcoma.
d. Hepatitis B virus - Hepatocellular carcinoma. (Vaccine is available).
e. Merkel polyoma virus-carcinoma of merkel cells of skin.

Outline of laboratory diagnosis of viral diseases:

1. Microscopic examination.
2. Isolation and identification of Virus.
3. Detection of viral antigen.
4. Detection of antibody.
5. Molecular diagnostic procedures.

For early diagnosis of infection, antigen detection and nucleic acid assay are most used.
Antibody appears at least after 7 to 10 days. Electron microscopy is very expensive, and

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Isolation and identification are time consuming, and technically demanding. So, culture
and electron microscope are not routinely used for diagnosis.

Microscopic examination:
1. Light microscopy can detect inclusion bodies, giant cells.
Inclusion bodies are viral aggregate or protein found in specific cells in specific
location and help in diagnosis. Inclusion bodies may be intracytoplasmic,
intranuclear or both. E.g., Negri bodies of rabies.

2. Fluorescent microscopy is used to detect viral antigen in specimen or tissue culture

by using fluorescein dye.
3. Electron microscopes can detect virus particle directly in specimen or in tissue
culture

Isolation and identification of Virus: Viruses cannot be grown on artificial culture

media. They are cultivated in tissue cultures, embryonated egg or by animal inoculation.
Virus growth in tissue culture is identified by varieties of ways. Some are given below.
a) By observing cytopathic effect: CPE is change in appearance of infected cell in cell
culture. CPE may produce change in size and shape of the cell, presence of giant cell,
inclusion bodies etc.
b) Interference
c) Hemadsorption.
d) Direct immunofluorescence assay
e) Electron microscopy

Detection of viral antigens: Antigens can be detected in serum or other specimens by

procedures such as ELISA, immunochromatographic tests, Direct immunofluorescence.

Detection of antiviral antibody: It is the most common method used to diagnose a

case. Appearance of IgM or four-fold rise of IgG indicates recent infection. Presence of
IgG (without recent rise) indicate past infection. ELISA, ICT, Western blot techniques are
commonly used to detect antibody.

Molecular diagnostic procedures such as PCR, RT-PCR, and many others detect viral
nucleic acid in patient’s specimen.

Antiviral agents.
1. Attachment blocker- neutralizing antibody.
2. Inhibitor of viral penetration- fuzeon(HIV).
3. Inhibitor of uncoating- amantidine (Influenza virus).
4. Translation inhibitor- interferon
5. Inhibitor of genome replication- acyclovir (herpes virus), lamivudine (HIV)
6. Inhibitor of viral release-oseltamivir (influenza virus).

Interferons are host-coded proteins, member of cytokine family which can inhibit viral
replication and have many immunomodulating function.
Interferons are produced in response to-viral infection, bacterial endotoxin, and various
antigens.

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Types:
1.  interferon.
2.  interferon.
3.  interferon.

 and  interferons are produced by many cell types in response to viral infection.
 interferons are produced by Th cell in response to various antigens.

Functions:
• Antiviral activity:  and  INF has stronger anti-viral activity than  INF.
Interferons inhibit viral replication by preventing viral protein synthesis
(degrade viral mRNA).
• Immunomodulating activity:  INF can activate macrophage.

Therapeutic use:
INF- in the treatment of chronic active hepatitis by HBV and HCV.
INF- in multiple sclerosis.
INF--chronic granulomatous disease.