Viruses may be defined as acellular organisms whose genomes consist of nucleic acid( either DNA or RNA).

Obligately replicate inside host cells using host metabolic machinery and ribosomes.

They form a pool of components which assemble into particles called VIRIONS, which serve to protect the genome and to transfer it to other cells

Viruses are inert (nucleoprotein ) filterable Agents Are obligate intracellular parasites Cannot make energy or proteins independent of a host cell Viral genome are RNA or DNA but not both.

Viruses have a naked capsid or envelope with attached

proteins

Do not have the genetic capability to multiply by binary fission.

Viruses are non-living entities

A small virus has a diameter of about 20nm.

Parvovirus

A large virus have a diameter of up to 300nm. Poxviruses

Spherical Rod-shaped Brick-shaped Tadpole-shaped Bullet-shaped Filament

Shapes of Viruses :Rod-shaped( TMV)

Brick-shaped (Pox virus) .

Bullet-shaped (Rabies virus)

Shapes of Viruses

:Filament

Virion
The

complete infectious unit of virus

particle
Structurally

mature, extracellular virus

particles.

envelope

Capsid

Viral core

Viral core
The viral nucleic acid genome, In the

center of the virion, : Control the viral

heredity and variation, responsible for

the infectivity.

VIRUS GENOMES
DNA RNA + or -

The genome of a virus can be either DNA or RNA

Single Stranded Double Stranded

DNA-double stranded (ds): linear or circular

Segmented

Single stranded (ss) : linear or circular

RNA- ss: segmented or non-segmented Segmented Double Stranded

ss: polarity+(sense) or polarity (non-sense) Circular

ds: linear (only reovirus family)

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. Share the antigenicity

 The

core of a virus particle consisting of the of proteins = Structural proteins

genome plus a complex of proteins.

complex

+Non- Structural proteins (Enzymes

&Nucleic acid binding proteins)

Helical Cubic /Icosahedral Complex

Helical symmetry

Cubic or icosahedral symmetry

 A well

known example is the tailed

bacteriophages such as T4.

The

head of these viruses is cubic with a

triangulation number of 7. This is attached by a collar to a contractile tail with helical symmetry.

A lipid-containing membrane that surrounds some viral

particles.

It is acquired during viral maturation by a budding process through a cellular membrane, Viruses-encoded glycoproteins are exposed on the surface of the envelope.

Not all viruses have the envelope, and viruses can be divided into 2 kinds: enveloped virus and naked virus.

Antigenicity some viruses possess neuraminidase

Infectivity Resistance

Different steps include

1. Adsorption
2. Penetration

3. Uncoating
4. Biosynthesis 5. Maturation 6. Release

Adsorption of a Naked Virus to a Susceptible Host Cell

To initiate infection cycle, a virus must first recognize and bind to a suitable host cell.
High specificity characterizes the interaction between virus and host. Receptors specific cell surface component of the host to which the virion attaches (ex. Proteins, carbohydrates, glycoprotein, lipids and lipoproteins)

Adsorption of an Enveloped Virus to a Susceptible Host Cell

The virion enters the intracelullar environment. Pinocytosis (viropexis) Engulfment of the virus particle by the plasma memrane and the subsequent production of an intracellular membranebound vesicle containing the virus particle Fusion of the viral envelop with the host cell membrane Not only does this method internalize the virus, itt can lead to fusion beween this and the other host cells nearby, forming multinucleated cells called syncytia.

Penetration of an Enveloped Virus by Fusion of Its Envelope with the Host Cell Membrane

Penetration of a Naked Virus by Rearrangement of Capsid Proteins

Removal of capsid from the virion Necessary to release viral genome before the viral DNA or RNA is delivered to its intracellular site of replication

Production of virally encoded proteins and replication of viral genome Early in infection thevirus redircts cell metabolism to synthesize new viral nucleic acid and proteins. Late in infection, structural proteins that are subunits of the virus coat are synthesized.

Construction of new nucleocapsids Assembly of structural subunits (and membrane components in enveloped viruses) and packaging of nucleic acid into new virus particles.

Direct release Lysis - mature virions or new infective viruses reach the extracelullar space, killing the host cell in some cases Budding - budding may also occur with or without cell death

From the stage of penetration, till the

appearance of mature daughter virions,

the virus cannot be demonstrated inside

the host cell. This period during which the virus seems to disappear or go underground is known as the eclipse phase.

VIRION DEFECTIVE VIRUS ABORTIVE INFECTION

Virus infection in some cells does not lead to production of infectious progeny. In such

cells, the viral components may be

synthesized but maturation or assembly is defective, and either no release occurs or the progeny is non infectious. This is known as abortive infection.

Viruses which are genetically deficient

and therefore incapable of producing infectious daughter virions without the helper activity of another virus are known as defective viruses. Eg, Hepatitis D virus and adeno associated satellite viruses.

Originally observed with influenza virus by Hirst.

Hemagglutination is due to the presence of

hemagglutinin spikes on the surface of the

virus.

Influenza virus also carries on its surface another peplomer, the enzyme neuraminidase which acts on the receptor & destroys it.

Neuraminidase is also called the receptor destroying enzyme (RDE).

Destruction of the receptor leads to the

reversal of hemagglutination and the

release of the virus from the red cell

surface. This is known as elution.

Elution is found only in myxoviruses that possess neuraminidase. With other viruses, hemagglutination is stable.

Used for detection and assay of virus.

Red cells which are not agglutinated, settle at the bottom in the form of a button, while the agglutinated cells are seen spread into a shield like pattern. As haemagglutinaion is specifically inhibited by the antibody to the virus,

haemagglutination inhibition provides a

convenient test for the antiviral antibody.

3 methods employed are

1.

Animal inoculation

2.
3.

Embryonated eggs
Cell cultures

Disadvantages Cost Maintenance Interference of immune system Individual variations Difficulty in choosing of animals for particular virus

Various routes of inoculation a)Yolk sac

b) Allantoic sac
c) Chorioallantoic membrane d. Amniotic cavity e. Intravenous

3 types of tissue cultures are available

1. Organ culture Eg, tracheal ring organ

culture for corona virus.

2. Explant culture Eg, adenoid tissue explant

culture for adeno virus

3. Cell culture

In cell culture, tissues are dissociated into

the component cells by proteolytic enzymes. The cells are washed, counted & suspended in a growth medium. The cell suspension is dispensed in

bottles, tubes or petri dishes. The cells

adhere to the glass surface and on incubation, divide to form a confluent

monolayer sheet of cells covering the

surface within about a week.

Cell cultures are classified into 3 types,

1. Primary cell cultures Eg, monkey kidney,

human embryonic kidney, human amnion & chick embryo.

2. Diploid cell strains Eg, WI -38, HL 8

3. Continuous cell lines Eg, HeLa, Hep -2, Vero

etc

1. 2.

Cytopathic effect (CPE) Metabolic inhibition

3.
4.

Hemadsorption
Interference

5.
6.

Transformation
Immunofluorescence

Cytopathic effect of enterovirus 71 and HSV in cell culture: note the ballooning of cells. (Virology Laboratory, Yale-New Haven Hospital, Linda
Stannard, University of Cape Town)

Syncytium formation in cell culture caused by RSV (top), and measles virus (bottom).
(courtesy of Linda Stannard, University of Cape Town, S.A.)

Syncytial formation caused by mumps virus and haemadsorption of erythrocytes onto the surface of the cell sheet.

Virus can be assayed in 2 ways:

1. Assay of total viral particles 2. Assay of infectious virions only.

Total particle assay can be performed by

Electron microscope Hemagglutination

Assay of infectivity

Plaque assay
Quantitative assay

Pock assay

Qualitative assay

Viruses can be classified into two ways: Based on the disease they caused or on the place of their isolation. The Universal System of Virus Taxonomy

Viruses are classified into 2 main

divisions depending on the type of

nucleic acid they possess:
1. Riboviruses
2. Deoxyriboviruses

 Viroids

are small, circular RNA

molecules with a rod-like secondary structure which possess no capsid or

envelope which are associated with

certain plant diseases.

Prions are rather ill-defined infectious agents

believed to consist of a single type of protein

molecule with no nucleic acid component.

These agents are associated with diseases such

as Creutzfeldt-Jakob disease in humans,

scrapie in sheep & bovine spongiform

encephalopathy (BSE) in cattle.

 Viral infection may cause a broad spectrum

of effects, ranging from no apparent cellular

damage to rapid cell destruction.

Polio virus cell death Molluscum contagiosum cellular

proliferation
Oncogenic viruses cellular proliferation

Steady state infection peaceful

coexistence

In tissue cultures cellular changes

(CPE)

Appearance of inclusion bodies

Structures with distinct size, shape, location & staining properties that can be demonstrated in virus infected cells under light microscope.

They are generally acidophilic(can be seen as pink structures when stained with

Giemsa/Eosin stain)

Helps in diagnosis

Cytoplasm Pox viruses

Nucleus Herpes viruses

Both Measles virus

Inclusions may be crystalline aggregates

of virions or made up of virus antigens

present at the site of virus synthesis.

Some inclusions represent degenerative

changes produced by viral infection.

Intracytoplasmic eosinophilic inclusions Negri bodies

Guarnieri bodies Vaccinia virus

Cowdry type A

Intranuclear inclusions
Cowdry type B

1. Inapparent (subclinical) infection 2. Apparent (clinical/ overt) infection

i. Acute

ii. Subacute iii. Chronic

3. Latent infection

1. 2.

Respiratory tract Alimentary tract

3.
4.

Skin
Conjunctiva

5.

Genital tract

Immunological response Mechanism Non- specific response

1.

Phagocytosis macrophages have more role than PMNLs

2.

Body temperature exception is fever

blisters.

3.

Hormones corticosteroids enhances viral infection.

4. Malnutrition increased complication

5. Age both extremes of age susceptible

6. Interferons
Family of host coded proteins produced by

cells on induction by viral or non viral

inducers.
IFN by itself has no direct action on viruses

but acts on other cells of the same species, rendering them refractory to viral infection.

 On exposure to interferon, cells produce a

protein (TIP), which selectively inhibits translation of viral mRNA, without affecting

cellular mRNA.
TIP block translation of viral mRNA into viral

proteins.
1. Protein kinase

2. Oligonucletide synthetase 3. RNAase

IFN are species specific.

Not virus specific. RNA viruses are better inducers than DNA viruses.

Production is increased by increasing temp. upto about 40 C

Inhibited by steroids and oxygen tension.

Based on antigenic characters, cell of origin and other properties, classified inot 3

types.
1. 2.

Alpha interferon (IFN- )- leukocytes Beta interferon (IFN )- fibroblasts, epithelial cells Gamma interferon (IFN ) T

3.

lymphocytes

Antiviral effects Antimicrobial effects Cellular effects Immunoregulatory effects

 1. 2. 3.

Diagnostic methods include Virus isolation & identification Direct detection Serodiagnosis

1. 2. 3.

Cell cultures Animal inoculation Embryonated eggs

1. 2. 3. 4.

Electron microscope Histophathology/ cytopathology Immunofluorescence/ immunochemistry Nucleic acid detection

1. 2. 3. 4. 5.

CFT HI Solid phase immunoassay Neutralization test Immune adherence hemagglutination

6.
7. 8.

Immunofluorescence assays
Immunoblot assays Particle agglutination assays

Viral vaccines Live viral vaccines Eg; Small pox

vaccine, measels

Killed vaccine Eg; Rabies vaccine,