Introduction To Virus

INTRODUCTION TO
VIRUS
INTRODUCTION
• A Virus is a submicroscopic infectious agent that replicates only inside
the living cells.
• Viruses are the smallest unicellular organisms that are obligate
intracellular.
• Viruses infect all humans, animals, plants & bacteria.
• Viruses are the most primitive microorganisms infecting man.
• Virus was discovered by Dmitri Ivanovsky in 1892.
• The study of viruses is known as virology.
• They do not have a proper cellular organization.
Difference between bacteria and virus
BACTERIA VIRUS
Cellular organisation + -
Growth in inanimate media + -
Binary fission + -
Both DNA & RNA + -
Ribosomes + -
Sensitivity to antibacterial antibiotics + -
Sensitivity to interferon - +
CHARACTERISTIC FEATURES
• They are obligate intracellular
• They possess either DNA or RNA, but never both.
• Filterable: They are smaller than bacteria, can be passed through the
bacterial filters.
• They cannot be grown on artificial cell free media (However, grow in
animals, eggs or tissue culture).
• They multiply by a complex method, but not by binary fission as seen in
bacteria.
• They do not have a proper cellular organization. They do not have cell wall
or cell membrane or cellular organelles including ribosomes, mitochondria.
• They lack the enzymes necessary for protein and nucleic acid synthesis.
• They are not susceptible to antibacterial antibiotics
MORPHOLOGY
Size of Viruses
• Viruses are extremely small, vary from 20–400 nm in size
• Largest virus: Pox virus (300 nm)
• Smallest virus: Parvovirus (20 nm)
Shape of Viruses
Most of the viruses are roughly spherical except:
Symmetry of Virus
• Icosahedral (cubical) symmetry: All DNA viruses (except poxviruses)
and most of the RNA viruses have icosahedral symmetry
• Helical symmetry: The capsomeres are coiled surrounding the nucleic
acid in the form of a helix or spiral. Such viruses are often fexible.
Example: rhabdoviruses, floviruses
• Complex symmetry: Poxviruses do not have either of the above
symmetry, but they possess a complex symmetry
COMPONENTS OF VIRUS
The entire virus particle called as virion, comprises of a nucleic acid (DNA
or RNA) surrounded by a protein coat called as capsid, together known as
the nucleocapsid. Some viruses also have an outer envelope.
• Nucleic Acid : Viruses have only one type of nucleic acid, either DNA or
RNA but never both. Accordingly, they are classified as DNA viruses and
RNA viruses. The nucleic acid may be single or double stranded, circular
or linear, segmented or unsegmented.
• Most DNA viruses possess dsDNA, except parvoviruses, which have
ssDNA
• RNA viruses possess ssRNA, except: Reoviruses (e.g. rotaviurs)—possess
dsRNA & Retroviruses—possess two copies of ssRNA.
• Capsid : Capsid is composed of a number of repeated protein subunits
(polypeptides) called capsomeres. Functions of capsid include:
• It protects the nucleic acid core from the external environment, e.g.
nucleases
• In non-enveloped viruses, it initiates the first step of viral replication by
attaching to specific receptors on the host cells, thus facilitating the
entry of the virus
• It is antigenic and specific for each virus
• Envelope : Certain viruses possess an envelope surrounding the
nucleocapsid. Envelope is lipoprotein in nature.
• The lipid part is derived from the host cell membrane and the protein
part is virus coded, made up of subunits called peplomers, which
project as spikes on the surface of the envelope
• Some viruses may have more than one kind of peplomers, e.g.
influenza viruses possess hemagglutinin and neuraminidase peplomers
• Enveloped viruses are more susceptible to heat and lipid solvents like
ether
• Peplomers are antigenic. They can also bind to specific receptors on
the host cells, thus facilitating the entry of the virus
NOMENCLATURE AND
CLASSIFICATION
• International Committee on Taxonomy of Viruses (2000) had
proposed a classification for viruses
• Viruses are grouped into families (ending with the suffix ‘viridae’) on
the basis of morphology, genome structure, and strategies of
replication
• Viruses infecting humans belong to 24 families, out of which
important ones are listed below
• Most of the families are further classified into genera (ending with the
suffix –‘virus’) based on physicochemical or serological differences
DNA viruses
RNA viruses
VIRAL REPLICATION
• Viruses do not undergo binary fission (seen in bacteria), but undergo a
complex way of cell division. Replication of viruses passes through six
sequential steps:
1. Adsorption/attachment
2. Penetration
3. Uncoating
4. Biosynthesis
5. Assembly
6. Maturation
7. Release of daughter virions
Diagrammatic representation of viral replication
1. Adsorption/attachment is the first and the most specific step of viral
replication. It involves receptor interactions between virus and host.
2. Penetration: After attachment, the virus particles penetrate into the host
cells either by
○ Phagocytosis (or viropexis)-Through receptor mediated endocytosis
○ Membrane fusion: seen in HIV
○ Injection of nucleic acid: seen in bacteriophages
3. Uncoating: Capsid is lysed (due to host lysozymes) and the nucleic acid is
released. This step is absent for bacteriophages.
4. Biosynthesis :In this step, the following viral components are
synthesized:
Nucleic acid
Capsid protein
Enzymes required for various stages of viral replication
Regulatory proteins to shut down the host cell metabolism.
• Site of Nucleic acid replication

• In DNA viruses, the DNA replication occurs in the nucleus; except in
poxviruses (cytoplasm).
• In RNA viruses: replication occurs in cytoplasm; except in retroviruses
and orthomyxoviruses (nucleus).
5. Assembly: Viral nucleic acid and proteins are packaged together to form
progeny viruses (nucleocapsids).
• DNA viruses are assembled in the nucleus except hepadnaviruses and
poxviruses (in cytoplasm)
• RNA viruses are assembled in the cytoplasm.
6. Maturation: Take place either in the nucleus or cytoplasm or membranes
7. Release of daughter virions occur either by:

• Lysis of the host cells as shown by nonenveloped viruses and
bacteriophages. OR
• Budding through host cell membrane as shown by enveloped viruses.
PATHOGENESIS OF VIRAL
INFECTIONS
Most of the viral infections progress through the following steps inside
the human body:
• Transmission (entry into the body)
• Primary site replication
• Spread to secondary site
• Manifestations of the disease.
LABORATORY DIAGNOSIS OF VIRAL
INFECTIONS
Laboratory diagnosis of viral infections is useful for the following purposes:
• To start antiviral drugs for those viral infections for which specific drugs are
available such as herpes, CMV, HIV, influenza and respiratory syncytial virus
(RSV)
• Screening of blood donors for HIV, hepatitis B and hepatitis C helps in the
prevention of transfusion transmitted infections
• For outbreak or epidemic investigation, e.g. influenza epidemics, dengue
outbreaks—to initiate appropriate control measures
• To start (PEP) post-exposure prophylaxis of antiretroviral drugs to the health
care workers following needle stick injury
• Surveillance purpose: To assess the disease burden in the community by
estimating the prevalence and incidence of viral infections
Laboratory diagnosis of viral infections can be
done by following methods:
1. Direct Demonstration of Virus

2. Detection of Viral Antigens
3. Detection of Specific Antibodies
4. Molecular Methods to Detect Viral Genes
5. Isolation of Virus
1. Direct Demonstration of Virus
• Electron microscopy: Detection of viruses by electron microscopy (EM) is
increasingly used nowadays. Specimens are negatively stained by
potassium phosphotungstate and scanned under EM.
• Immunoelectron microscopy: The sensitivity and specificity of EM can be
improved by adding specific antiviral antibody to the specimen to
aggregate the virus particles which can be centrifuged. The sediment is
negatively stained and viewed under EM.
• Fluorescent microscopy: Direct immunofluorescence (Direct-IF) technique
is employed to detect viral particles in the clinical samples.
• Light microscopy: To demonstrate inclusion bodies
2. Detection of Viral Antigens : Various formats are available for the
detection of viral antigens in serum and other samples such as enzyme-
linked immunosorbent assay (ELISA), immunochromatographic test (ICT),
flow through assays, enzyme-linked fluorescence assay (ELFA), etc.
HBsAg and HBeAg antigen detection for hepatitis B virus infection from
serum
NS1 antigen detection for dengue virus infection from serum
SARS-CoV-2 antigen (nucleocapsid protein) detection in nasopharyngeal
swabs by immunochromatographic assay
p24 antigen detection for HIV infected patients from serum
3. Detection of Specific Antibodies
Antibody detection from serum is one of the most commonly used method
in diagnostic virology. Appearance of IgM antibody or a four-fold rise of
titer of IgG antibody indicates recent infection; whereas the presence of
IgG antibody (without a recent rise) indicates chronic or past infection.
• Techniques such as ELISA, ELFA, ICT, flow through assays are widely used
for antibody detection against most of the viral infections, for example:
Anti-HBc, Anti-HBs and Anti-HBe antibodies in serum for hepatitis B
infection
Anti-Hepatitis C antibodies in serum
Antibodies against HIV-1 and HIV-2 antigens from serum
Anti-Dengue IgM/IgG antibodies from serum
4. Molecular Methods to Detect Viral Genes
Advent of molecular techniques has eased the diagnosis of viral infections.
They are more sensitive, specific and yield quicker results than culture.
Different types of molecular methods:

Nucleic acid probe—for detection of DNA or RNA by hybridization
PCR—for DNA detection by amplification
Reverse transcriptase-PCR—for RNA detection
Real time PCR—for DNA quantification
Real time RT-PCR—for RNA quantification.
5. Isolation of Virus
Viruses cannot be grown on artificial culture media. They are cultivated by
Animal inoculation : Because of the ethical issues related to use of animals,

animal inoculation is largely restricted only for research purpose.
Embryonated egg inoculation : Embryonated eggs were first used for viral
cultivation by Good pasture in 1931. Subsequently, this technique was widely
used in the past.
Tissue cultures: Enders, Weller, and Robbins (1949) used tissue cultures of
non-neural origin to cultivate poliovirus and that was the turning point
following which tissue culture technique was widely used in diagnostic
virology.
3 types of tissue cultures : Organ culture, explant culture & cell line culture
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INTRODUCTION TO

• Site of Nucleic acid replication

6. Maturation: Take place either in the nucleus or cytoplasm or membranes

7. Release of daughter virions occur either by:

1. Direct Demonstration of Virus

Different types of molecular methods:

Animal inoculation : Because of the ethical issues related to use of animals,

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