Introduction to virology

Classification
Structure
Replication

DR.ARVIND M.KURHADE
M.B.B.S.,M.D.
Course convener MDSC 1002 Lecturer In
Clinical Microbiology F.M .S
; U .W . I .
 Objectives

1) Introduction
2) Classification
3) Morphology and Structure
4) Replication
5) Cultivation
6) Bacteriophage
 Introduction to Virology
• A virus is an Obligate
Intracellular parasite
containing genetic material
surrounded by protein. - Most
viruses range in sizes from 20 –
300 nanometers.
• Have a receptor - binding
protein for attaching to cells.
• Ultra microscopic , Filter
passing.[ Does not possess Both
DNA and RNA =Virus]
• Virus particles can be observed Viral genome is packaged in protein
coat
by an Electron Microscope.
 Introduction to Virology
• Classification of viruses is based on nucleic acid
• RNA viruses.
• DNA viruses.
• Replication
• Depends on host cell machinery for replication utilizing the
host cell’s nucleic acids.
• Cultivation of viruses is complex & includes three methods
• Animal Inoculation
• Embryonated Egg Culture
• Cell Culture
• Virus can not cultivated on Chemically defined media
or Artificial culture media which don’t contain Live
Cells.
Intro contd…

• Viruses : - Much smaller than bacteria . They contain DNA

or RNA . They are not readily destroyed by antibiotics .
The outer shell is composed of individual building blocks called
Capsomeres .

• “Filterable agents” – Can pass through filters that can hold back bacteria

• Vary widely in size:

– Largest – Poxvirus ( 300 nm )
– Smallest – Parvovirus ( 20 nm )

• Virion – Extracellular Infectious Virus Particle

 The Extracellular Infectious
Virion : Virus Particle [ DNA
covering material is called
= Capsid.] .
Envelope

Capsid

Viral core
 Shape of
Viruses
• Spherical-:Poliovirus

• Rod-shaped-:Tobacco
mosaic virus.

• Brick-shaped-:Pox
virus

• Bullet-shaped-:Rabies
virus
•VIRAL
CLASSIFICATION
D N A- OR- R N A
Classification of viruses- The International
Committee on Taxonomy of Viruses
Basis of classification :
1. Virion morphology : Size , Shape, Type of
Symmetry , Presence or Absence of
Peplomers .
2. Physicochemical properties : Molecular
mass , Density , pH & Thermal stability ,
Susceptibility to physical & chemical agents
especially ether & detergent.
Classification of viruses contd…
3. Virus genome properties : Type of nucleic acid ( DNA
or RNA ) , size of genome , number of strands ( single or
double) , linear or circular. ( VIRUS - does not possess
both DNA And RNA ) .

Classification by Symptomatology :
The oldest classification of viruses based on Disease
they produce --> convenient for Clinicians .
*Not satisfactory for the Biologists .
*Completely unrelated Viruses may produce similar
diseases ( e.g. Respiratory Infections ) .
 DNA VIRUSES

ENVELOPED NAKED capsid

Double – stranded Double – stranded Single –

stranded

Icosahedral Complex Icosahedral Icosahedral

HERPES POX PAPOVA PARVO

 RNA VIRUSES
Single – stranded Double – stranded

Positive stranded Negative stranded

Naked Enveloped Enveloped Naked

*PICORNA *TOGA BUNYA *REO

*CALICI *FLAVI ORTHOMYXO
CORONA PARAMYXO
RETRO RHABDO
ARENA

* Icosahedral ; all of the rest have Helical symmetry

•MORPHOLOGY
& STRUCTURE
Basic virus structure

From Medical Microbiology, 5th ed., Murray, Rosenthal & Pfaller, Mosby Inc., 2005, Fig. 6-1.
Morphology & virus structure
RNA
• Nucleus → Nucleic acid core (genome)
DNA
• Capsid : DNA covering material of VIRUS:.

Nucleus & Capsid → Nucleocapsid

• Geometric shapes –Nucleocapsid symmetry

• Helical symmetry
• Icosahedral pattern (Cubical symmetry )
• Complex

• Envelope - May be present or absent

• Two layers : Protein & Lipid
 Structures compared

From Medical Microbiology, 5th ed., Murray, Rosenthal & Pfaller, Mosby Inc., 2005, Fig. 6-4.
 Morphology – Structure & Shape of a virus
• Nucleic acid
• & Capsid with or without
Envelope.
• Capsid = DNA coviring
material of VIRUS = The
protein coat surrounding the
nucleic acid core. It
– Protects nucleic acid from
inactivation
– Helps to introduce viral
genome into host cell
• Capsomers - The outer shell is
composed of : repeating protein
subunits blocks that make up
the capsid.
• Protomers – The polypeptide
chains which make up the
capsomers
Structure & Shape of a virus : Capsid Symmetry
Icosahedral – A polygon with 12 Corners
(Vertices ) & 20 Sides ( Facets ) .
• Side – Equilateral triangle
• Two types of Capsomers form the
Capsid
• Pentagonal capsomers form the
vertices
• Hexagonal capsomers form the
Sides.
Helical – The capsomers & nucleic acid
are wound together to form a helical or
spiral tube.
Complex - The overall shape of virus is
quite variable , but mostly they are
spherical ( Pox-virus )
Dr Ekta, Microbiology
Morphology – Structure & Shape of a Virus -
Envelope :
• May or may not be present. Enveloped Icosahedral- HIV
• Derived from the host cell
membrane.
• Lipoprotein in nature – Lipid is
of host cell origin while protein
is from virus.
• Protein subunits seen as
projecting spikes on the
surface of envelope – called
Peplomer .
• A virus may have more than one
type of peplomer e.g. Influenza
virus.
• Confers chemical , antigenic &
biological properties. ENVELOPE
• Susceptible to lipid solvents
Peplomer
• VIRAL
REPLICATION
Viral Replication –A General Scheme
Stages in virus replication begin when virions infect cells.
1) Recognition of host cell
↓
2) Attachment
↓
3) Penetration
↓
4)Uncoating
↓
5) Transcription
↓
6) Protien synthesis
↓
7) Replication
↓
8) Assembly
↓
9) Lysis & release
ATTACHMENT

PENETRATION HOST
UNCOATING FUNCTIONS

Transcription
Translation
REPLICATION

VIRAL LIFE
ASSEMBLY
CYCLE (MATURATION)

RELEASE

MULTIPLICATION
 Viral Replication

• Eclipse phase – From the stage of penetration

till the appearance of mature daughter virions,
the virions cannot be detected inside the host
cell.
• The duration of eclipse phase or replication
time is about 3-12 hours for most of the
viruses.
•CULTIVATION of
viruses-can’t grow on
chemically defined media.
Cultivation of viruses – (1) ANIMAL
Type , age , method of inoculation
depend on type of viruses
Herpes simplex virus : Rabbit cornea Vesicles
Rabies virus : White Mouse (baby/adult) Intracerebral
1 – 3 weeks encephalitis/ rabies
Dengue virus : Baby White Mouse, 1 – 3 days old
Intracerebral/ subcutaneously
3 – 7 days tremor , paralysis
Polio virus : Monkey intracutaneously , IM ,
intraneural , intraspinal
paralysis
 Cultivation of virus- (2) Embryonated egg
Inoculation methods depend on the type of
viruses.
Chorioallantoic membrane (CAM) : 10 –
12 days old embryo
e.g.: Pox virus , Herpes simplex virus
↓ Pocks / Plaque
Amniotic cavity : 10 - 11 days old embryo
e.g :Influenza virus , Mumps virus.

Allantoic cavity : 10 days embryo

can be propagated in much larger quantities,
used for vaccine production eg:
Influenza , Rabies vaccine.

Yolk sac : 3 – 8 days old embryo –e.g.:

Chlamydiae & Ricketssia.

Intraembryonal : 8 – 10 days old embryo

e.g.: Japanese B encephalitis virus
Cultivation of virus (3) Cell culture

For isolation , Neutralization , Production of Antigen for serological

assay
3 basic type of cell culture :
Primary culture :- Made by dispersing cells from freshly removed
host tissues , unable to grow for more than a few passage. eg : -
Rhesus monkey kidney cell & Human amnion cell culture .
Secondary / Diploid culture :- Diploid cell lines up to 50 passage
but retain their normal chromosome pattern. eg Human embryonic
lung cell culture.
Continuous cell line / Heteroploid cell culture :- Capable of
prolonged , infinite growth , derived from cancerous cells , grow
faster with haploid chromosomes. eg HeLa cell from cancer of
cervix , Hep-2 ( epithelioma of larynx cell ) .
Virus isolated clinical samples by cultivation on -1) Animal 2)
Embryonated eggs & 3) Cell culture .
Not cultivated on chemically defined media.
Virus grows in cell culture & produces following effects:
1. Cytopathic effect uninfected

2. Hemadsorption

3. Interference

respiratory syncytial virus

4. Transformation

5. Immunofloroscence

6. Electron Microscopy
 Inclusion body formation
In the course of virus multiplication within cells → Virus
-specific structure called inclusion bodies may be
produced .They become far larger than the individual
virus particle with affinity for acid dyes ( eosin ) .
The inclusion bodies can be present in
a) Nucleus : Herpes virus , Poliovirus , Echovirus ,
Coxsackie virus , Influenza virus , Rubella virus , CMV .
b) Cytoplasm : Poxvirus , Rabies virus
c) Both ( nucleus&cytoplasm):Adeno virus & Measles virus

Negri body (Intra

cytoplasmic)- Rabies
 Inclusion bodies
Intranuclear & Intracytoplasmic , mostly
• Presumptive etiologic diagnosis acidophilic , Few basophilic , like
• Intra nuclear inclusions Adenovirus

( Mostly DNA viruses ) .

Cowdry type A ( Yellow fever )-
Variable size,granular appearance
Cowdry type B ( Polio)
-Circumscribed , multiple
• Intra cytoplasmic inclusions (
Mostly RNA viruses)
Guarnieri bodies ( Small pox )
Negri bodies ( Rabies )
Both ( Measles , Adenovirus )

Owl eye (Intranuclear )- CMV

 Major Diseases Important Viruses
AIDS HIV-1
HAV, HBV, HCV,
Hepatitis
HDV, HEV, CMV, EBV
HPV, HCV, EBV,
Cancer
HBV, HHV-8, HTLV-I
HSV, togaviruses, flaviviruses,
Encephalitis
rabies, picornaviruses
Enteroviruses, mumps,
Meningitis
HSV-2, VZV
Rotavirus, caliciviruses,
Gastrointestinal Tract
adenoviruses (40, 41)
Rhinoviruses, coronaviruses,
Common Cold
adenoviruses
Influenza, parainfluenza,
Lower Respiratory Tract
ReSV,adenoviruses, SARS-CoV
Congenital & Neonatal CMV, Parvovirus B19,
Infections rubella, HSV, HIV
 Bacteriophage
Viruses that infect bacteria.
• Capsid ( head ) is polyhedral ,
Sheath is helical.
• Tail fibers , plate and pin.
• High Host specificity.
• Pass through bacterial filters.

• Commonest habitat : - Intestinal

tract of animals .

Bacteriophage are mostly needed for

Bacterial identification .
 Importance of Bacteriophage - Mostly
needed for bacterial identification
• Act as carrier of genes from one
bacterium to another.

• Phage typing
• Sub typing of a species or genus
e.g. Salmonella , Vibrio.

• Utilized to study Host - Parasite

Relationships