VIROLOGY

DEFINITION OF VIRUSES:

Viruses are small non-cellular intracellular infectious agents that use a living cell for their
replication and have an extracellular state i.e., they cannot reproduce and carry on metabolic
processes without a host cell. Viruses are ultramicroscopic particles containing nucleic acid
(DNA or RNA) surrounded by protein shell called capsid.

Size of viruses: The size of viruses ranges from 20 nm to 300 nm in diameter, the largest
being the small pox virus and the smallest being the polio virus.

Virions: While not inside an infected cell or in the process of infecting a cell, viruses exist in
the form of independent particles. These viral particles are known as virions.

STRUCTURE OF VIRUSES:

All viruses contain the following components: 1) a nucleic acid genome and 2) a protein
capsid that covers the genome. Together this is called the nucleocapsid. In addition, many
animal viruses contain 3) Envelope.

1) Nucleic acid: Viruses may contain ribonucleic acid (RNA) or deoxyribonucleic acid
(DNA). The nucleic acid portion of the viruses is known as the genome. The nucleic acid
may be single-stranded or double-stranded; it may be linear or a closed loop.

2) Capsid: The genome of the virus is surrounded by a protein coat known as a capsid,
which is formed from a number of individual protein molecules
called capsomeres. Capsomeres are arranged in a precise and highly repetitive pattern around
the nucleic acid. The combination of genome and capsid is called the viral nucleocapsid.
Function of capsid: Three important functions of capsid are:
a) Protects the nucleic acid from digestion by enzymes.
b) Contains special sites on its surface that allows the virus particle to attach to the host cell.
c) Provides proteins that enable the virus particle to penetrate the host cell membrane.

3) Envelopes: Some kinds of viruses contain envelopes. The envelope of viruses is a lipid
bilayer containing glycoproteins embedded in the lipid. The envelope gives a somewhat
circular appearance to the virus. Projections from the envelope are known as spikes. The
spikes sometimes contain essential elements for attachment of the virus to the host cell. The
virus of AIDS, the human immunodeficiency virus, uses its spikes for this purpose.
Figure: General structure of a virus
CLASSIFICATION OF VIRUSES:

Viruses are broadly classified on the basis of the following five characteristics:

A) Classification on the basis of nucleic acid:

DNA virus

Double stranded Single stranded

Enveloped Unenveloped Unenveloped

(eg. Herpes, Pox) (eg. Adeno, papilloma)
(Eg. Parvovirus)

RNA virus

Double stranded (eg. Reovirus) Single stranded

+ RNA - RNA
(eg. Polio, influenza) (eg. Rabies)

1. DNA virus:

 Viral genome is DNA

i) Double stranded DNA virus: eg. Adenovirus, Herpesvirus

ii) Single stranded DNA virus: eg. Parvovirus, φ174 virus

2. RNA virus:

 Viral genome is RNA

i) Double stranded RNA virus: eg. Reo virus

ii) Single stranded RNA virus: these are further classified into two groups
  Positive sense RNA (+RNA): Polio virus, Influenza virus
 Negative sense RNA (-RNA): Rabies virus

B) Classification on the basis of structure or symmetry

1. Cubical virus:

 They are also known as icosahedral symmetry virus.

 Eg. Reo virus, Picorna virus

2. Spiral virus:

 they are also known as helical symmetry virus

 Eg. Paramyxovirus, orthomyxovirus

3. Radial symmetry virus:

 eg.Bacteriophage

4. Complex virus:

 eg. Pox virus

C) Classification on the basis of replication properties and site of replication

1. Replication and assembly in cytoplasm of host:

 Eg. All RNA virus replicate and assemble in cytoplasm of host cell except Influenza
virus.

2. Replication in nucleus and assembly in cytoplasm of host:

 Eg. Influenza virus, Pox virus

3. Replication and assembly in nucleus of host:

 All DNA viruses replicate and assemble in nucleus of host cell except Pox virus.
D) Classification on the basis of host range:

1. Bacteriophage:
 Bacteriophage are virus infecting bacteria. Eg, T2, T4.

2. Plant virus:
 Those virus that infects plants. Eg. TMV, cauliflower mosaic virus
3. Animal virus:
 Those virus that infects animals. Eg. Polio virus, Retro virus, Herpes virus, Adeno
virus
4. Insect virus:
 Virus that infects insects. Eg. Entomopox virus, Granulosis virus

E) Classification on the basis of mode of transmission:

1. Virus transmitted through respiratory route:

 Eg, Swine flu, Rhino virus

2. Virus transmitted through faeco-oral route:

 Eg. Hepatitis A virus, Polio virus, Rota virus

3. Sexually transmitted viruses:

 Eg. Retro virus

4. Virus transmitted through blood transfusion:

 Eg. Hepatitis B virus, HIV

5. Zoonotic virus:

 Virus transmitted through biting of infected animals;

 Eg. Rabies virus, Alpha virus, Flavi virus
 PATHOGENESIS OF VIRAL INFECTION

Pathogenesis is the process or mechanism by which a viral infection leads to disease.

Pathogenic mechanisms of viral disease include (1) implantation of virus at the portal of
entry, (2) viral replication and (3) shedding of virus into the environment by spreading to
target organs (disease sites).

 Implantation at Portal of Entry: At the initial stage, the virus implants itself at the
entry portal into the body. Implantation is the earliest stage of pathogenesis. Viruses
usually implant on cells of respiratory, gastrointestinal, skin and genital tissues.
 Viral Replication and Spread: Viral replication and spread of the virus follows
implantation. The invading virus reproduces itself in large numbers. It usually does
this intracellularly. Replicated virus from the initially infected cell has the capability to
disperse to neighbouring extracellular fluids or cells. The most common route of
spreading of the virus is the circulatory system or through the blood stream. The
replicated virus then spreads to target organs (disease sites) throughout the body. Some
viruses such as Herpes, rabies and polio viruses, can also disseminate via nerves.
 Shedding of virus: The viruses then spread to sites where shedding into the environment
of the target sites can occur. The respiratory, alimentary and urogenital tracts and the
blood are the most frequent sites of shedding.

Diagram showing pathogenesis and replication of a DNA virus.

IMMUNOPROPHYLAXIS AGAINST VIRAL DISEASES:

Immunoprophylaxis against viral illnesses includes the use of vaccines or antibody-

containing preparations to provide immune protection against a specific disease. There are
two types of immunoprophylaxis.

1) Active Prophylaxis (Vaccines)

Active immunization involves administering a virus preparation that stimulates the body's
immune system to produce its own specific immunity. Viral vaccines available for use
include the following types:

(a) Attenuated live viral vaccines- Most live vaccines contain viruses that have been
attenuated by laboratory manipulation. These attenuated viruses can infect and replicate in
the recipient and produce a protective immune response without causing disease. Live
attenuated viral vaccines can often confer lifelong immunity after one immunization series.

(b) Killed (inactivated) viral vaccines- Killed viral vaccines contain either whole virus
particles, inactivated by chemical or physical means, or some component(s) of the virus.
Completely inactivated viral vaccines cannot cause infection. However, they do not generally
produce lifelong immunity following one immunization series.

(c) Recombinant produced antigens- Application of a recombinant DNA strategy to

develop new vaccines is performed by identifying the specific component(s) that can induce
the production of protective antibodies, and then cloning and expressing the gene encoding
that protein. Eg. Production of a safe and effective recombinant vaccine against hepatitis B
virus.

2) Passive Prophylaxis

Passive immunity is conferred by administering antibodies formed in another host. Human

immunoglobulins remain a mainstay of passive prophylaxis for viral illnesses; they are
usually used to protect individuals who have been exposed to a disease and cannot be
protected by vaccination. For eg. Rabies antidote.
CONTROL OF VIRAL INFECTIONS AND DISEASES

Virus infection can be controlled by several approaches such as improved sanitation, removal
and control of vectors responsible for disease and by use of antiviral chemotherapeutic
agents.

a) Improved sanitation: Many viral diseases are controlled by reducing exposure to the
virus by eliminating nonhuman reservoirs, improving sanitation and maintaining proper
hygiene. For eg., blood bank testing, for hepatitis B surface antigen and for antibodies to
HIV-1, HIV-2 and hepatitis C also avoids exposure by identifying and discarding blood units
contaminated with these infectious agents.

b) Control of vectors and non-human viral reservoirs: Control of transmission of viral

diseases is also achieved by elimination of vectors and non-human viral reservoirs which
carry the virus. Control of non-human viral reservoirs involves removal of stray dogs for
elimination of rabies diseases, quarantine of incoming pets and vaccination of domestic
animals. Removal of vectors such as mosquitoes responsible for yellow fever or dengue
disease is another approach for controlling of vector-borne viral diseases.

c) Antiviral chemotherapeutic agents: Antiviral chemotherapeutic agents can be divided

into three categories: virucidal agents, antiviral agents and immunomodulators.

Virucidal agents directly inactivate intact viruses. Eg. Topical treatment of warts with
podophyllin, which destroys both virus and host tissues. Antiviral agents inhibit viral
replication at the cellular level, interrupting one or more steps in the life cycle of the virus.
These agents have a limited spectrum of activity and, because most of them also interrupt
host cell function, they are toxic to various degrees. Immunomodulators are chemical
agents that have relatively specific noncytotoxic effects on the immune system. They
modify the immune response or the functioning of the immune system by the
stimulation of antibody formation or the inhibition of white blood cell activity .

LABORATORY DIAGNOSIS OF VIRAL DISEASES

In general, diagnostic tests for viral diseases can be grouped into 4 categories: (1) direct
detection, (2) indirect examination (virus isolation), (3) serology and (4) nucleic acid
detection.
1) Direct detection: Direct examination methods are also called rapid diagnostic methods
because they can usually give a result either within the same or the next day. In direct
examination, the clinical specimen is examined directly for the presence of virus particles,
virus antigen or viral nucleic acids. Virus particles are detected and identified on the basis of
morphology by electron microscope or histological examination via light microscope.

2) Indirect examination: In indirect examination, the specimen is inoculated into cell

culture, eggs or animals in an attempt to grow and detect the virus, this is called virus
isolation and identification.

3) Serology: In general, the majority of common viral infections can be diagnosed by

serology. A serological diagnosis can be made by the detection of rising titres of antibody
between acute and convalescent stages of infection, or the detection of IgM
antibody. Common techniques used for serological diagnosis are Radioimmunoassay (RIA),
Enzyme linked immunosorbent assay (EIA), Western Blot etc.

4) Nucleic acid detection: Methods based on the detection of viral genome are commonly
known as molecular methods. This technique allows the quantification of DNA/RNA present
in the specimen. Newer molecular techniques such as the polymerase chain reaction (PCR)
are used which allows extremely sensitive detection of infection.

LIST OF SOME COMMONLY USED ANTIVIRAL AGENTS:

Antiviral agent Uses

1) Acyclovir Genital HSV infections,

Mucocutaneous infections, chicken pox

2) Amantadine Influenza A prophylaxis and treatment

3) Adenine arabinoside HSV encephalitis
4) Didanosine HIV infection
5) Famciclovir Varicella-Zoster (shingles)
6) Foscarnet CMV retinitis in AIDS patients
7) Idoxuridine HSV keratitis
8) Interferon alpha Chronic Hepatitis B and C
9) Ribavirin RSV bronchitis/pneumonia
10) Zidovudine HIV infection prophylaxis and treatment