Virology

Virology

Dr. Shujaat Ali (RPh, PhD)

Assistant Professor
Dr. Shujaat Ali (RPh, PhD)
Assistant Professor
Introduction:
 Virus is a Latin word derived from Venom which means
“Poison”.

 Small infectious agent which caused infection in humans,

animals, plants and bacteria.

 Viruses contain DNA or RNA and a protein coat (Capsid

with Capsomeres) that protects the genetic material.

 Some are enclosed by an envelope

 Minuscule, Acellular.

 Obligate intracellular parasites (need living host cells to

replicate)
Introduction:
 Cannot carry out any metabolic pathway (No ATP generation
system, No ribosomes for protein systhesis)

 Neither grow nor respond to the environment.

 Cannot reproduce independently

 Some viruses have spikes

 Most viruses infect only specific types of cells in one host

 The Discovery of Virus:
 Edward Jenner first introduced the term “Virus” to
microbiology.

 Virus was first discovered by the Russian microbiologist

“Dmitri Ivanosky” in 1890 and named “Tobacco Mosaic
Virus”.

 In 1935 Wendell Stanley crystallized tobacco mosaic virus

(TMV); an accomplishment for which he was awarded a
share of the 1946 Nobel Prize in Chemistry
The Discovery of Virus:
Since Dmitri Ivanovsky's 1892 discovered that the agent which caused tobacco
mosaic disease was filterable. He obtained bacteria free filterate from ground up
infected plants and placed on the healthy leaves of tobacco. He found that
filterate produced the disease in healthy plant. After that the presence of similar
filter passing , ultramicroscopic agents was seen in the victims of many disease
including foot and mouth disease ( herpes and voricella infections) and yellow
fever as well. In 1935 stanley then purified these filterable agent by
crystallization and found that causative agent have only nucleic acid and protein
in their structure. Hence the agents are simply described as “virus”
Structure of Virus
Size of a Virus:
 Viral size is determined by Electron Microscopy.

 Most of the viruses are smaller than bacteria but larger

viruses (e.g. vaccinia virus) are same in size to very small
bacteria (mycoplasmas, rickettsias, chalamydias)

 Viruses range from 20nm-1000nm in length.

Size of a Virus:

“Microbiology, an Introduction” by tortora

Structure of a Virus:
 A Virion is a complete, fully developed, infectious viral
particle composed of nucleic acid and surrounded by a protein
coat outside a host cell.

 Viruses are classified by their nucleic acid and by difference

in the structures of their coat.

 Thus, viral nucleic acids can be DNA or RNA, double-

stranded or single-stranded, monopartite or multipartite, linear
or circular, as short as 2 kb or up to 2500 kb long.
Capsid of a Virus:
 The nucleic acid of virus is protected by a protein coat called
the capsid

 The structure of capsid is determined by the nucleic acid and

the mass of virus Capsid is composed of protein subunits
called capsomeres

 Capsomere may be composed of a single or several types of

proteins

 The arrangement of capsomere is characteristic of particular

type of virus
Envelope of a Virus:
 In some viruses, capsid is covered by an envelope consisting of
combination of lipids, proteins and carbohydrates.

 Viruses that lack enveloped are known as non-enveloped or naked viruses.

 All of the negative stranded RNA viruses are enveloped.

 It is acquired during the replication in the host cell and is unique to each
type of the virus.

 The envelop protects the nucleic acid from nuclease enzyme in biological
fluids and promotes attachment to host

 The structural components of envelope remains biologically active only in

aqueous medium and are destroyed in acidic medium. Therefore most of
the enveloped viruses are usually transmitted through body fluids (blood,
respiratory droplets, tissue exudates etc).
Envelope of a Virus:
 Spikes of a Virus:

 Viruses may or may not have spikes

 Spikes are carbohydrate-protein complexes that projects from the surface of
the envelope
 It helps attachment to the host cells.
 Spikes causes clumping of RBCs.
 Influenza virus
(haemaglutination)
 Spikes of a Virus:

 Viruses may or may not have spikes

 Spikes are carbohydrate-protein complexes that projects from the surface of
the envelope
 It helps attachment to the host cells.
 Spikes causes clumping of RBCs.
 Influenza virus
(haemaglutination)
General Morphology of Virus
General Morphology of Virus
 Viruses are classified on the basis of capsid
architecture

 Electron microscopy and X-Ray Crystallography

reveals the shapes of a capsid

 On the basis of Capsid structure:

1) Helical Viruses
2) Polyhedral Viruses
3) Enveloped Viruses
4) Complex Viruses
 Helical Virus:

 The viral genome is found within hollow cylindrical capsid that

has a helical structure.
 Rod like appearance
 May be rigid or flexible
 Examples are:
1) Rabies Virus
2) Ebola Virus
 Polyhedral Virus:

 The polyhedral virus appears as many sided appearance

 The virus consists of capsids in the shape of an icosahedron
(regular polyhedron with 20 triangular faces)
 The Capsomere of each face forms an equilateral triangle.
 Example=Adenovirus
 Enveloped Virus:

 The helical or polyhedral virus covered with envelope are called

enveloped viruses.
 Enveloped Helical Viruses i.e. Influenza Virus
 Enveloped Polyhedral Viruses i.e. Herpes Simplex Virus
 Enveloped Viruses are roughly spherical.
 Complex Virus:

 Bacterial Viruses
(Bacteriophage)
 These viruses possess a capsid
that is neither purely helical nor
purely icosahedral, and that may
possess extra structures such as
protein tails or a complex outer
wall.
 Complex Virus:

 They have a complex structure

consisting of an icosahedral head
bound to a helical tail, which may
have a hexagonal base plate with
protruding protein tail fibres.
 This tail structure acts like a
molecular syringe, attaching to the
bacterial host and then injecting the
viral genome into the cell.
Complex Virus:
 Taxonomy of a Virus:

• Viral species: A group of viruses sharing the same genetic

information and ecological niche (host).
• Thus Common names are used for viral species e.g. Human
Immunodeficiency Virus (HIV).
• Subspecies are designated by a number e.g. HIV-I
 The suffix–virus is used for genus names
 Family name ends in–viridae
 E.g. Family Herpesviridae, Genus Simplexvirus.
 Virus Families

 Families of Viruses that affect human are

1) Single-stranded DNA, Nonenveloped

viruses
• Parvoviridae
• Human parvovirus
• Fifth disease
• Anemia in immunocompromis ed patients
 Virus Families

2) Double-stranded DNA, Nonenveloped

viruses
• Adenoviridae
• Mastadenovirus
• Respiratory infections in human
• Tumors in animals
• Papovaviridae
• papillomavirus
• Cause tumours
 Virus Families
3) Double-stranded DNA, Enveloped viruses
• Poxviridae
• Orthropoxvirus
• Smallpox and cowpox

• Hepadnaviridae
• Hepadnavirus
• Hepatitis B, Liver tumor
 Virus Families
3) Double-stranded DNA, Enveloped viruses
• Herpesviridae
• Simplexvirus (HHV1 and HHV 2)
• Varicellavirus (HHV 3)
• Lymphocryptovirus (HHV 4)
• Cytomegalovirus (HHV 5)
• Roseolovirus (HHV 6)
• HHV 7
• Kaposi's sarcoma (HHV 8)

 Fever, Blisters, ChickenPox, Burkitt’s lymphoma

 Virus Families
4) Single-stranded RNA, + strand Nonenveloped
viruses
• Picornaviridae
• Enterovirus
• Rhinovirus (common cold)
• Hepatitis A virus
• Caliciviridae
• Hepatitis E virus
• Norovirus (Norwalk agent) causes
gastroenteritis.
 Virus Families
5) Single-stranded RNA, + strand Enveloped viruses
• Togaviridae
• Alphavirus
• Rubivirus (rubella virus)

• Flaviviridae
• Hepatitis C virus
• Flavivirus
• Arbovirus
 Virus Families
5) Single-stranded RNA, + strand Enveloped viruses
• Coronaviridae
• Coronavirus
• Upper respiratory tract infections
 Virus Families
6) Single-stranded RNA, - strand, one RNA Strand

• Rhabdoviridae
• Vesiculovirus
• Lyssavirus (rabies virus)

• Filoviridae
• Filovirus (Ebola Virus)
 Virus Families
6) Single-stranded RNA, - strand, one RNA Strand

• Paramyxoviridae
• Paramyxovirus (mumps virus)
• Morbillivirus (measles virus)

• Deltaviridae
• Hepatitis D virus
 Virus Families
7) Single-stranded RNA, - strand, multiple RNA Strands

• Orthomyxoviridae
• Influenzavirus (A, B and C)
• Enveloped Spikes can agglutinate RBCs.

• Bunyaviridae
• Hantavirus
• Haemorrhagic fever
 Virus Families
7) Single-stranded RNA, - strand, multiple RNA Strands

• Arenaviridae
• Arenavirus
• VEE (Venezuelan equine encephalitis ) and Lassa
Fever (viral hemorrhagic fever)
 Virus Families
8) Single-stranded RNA, produce DNA

• Retroviridae
• Lentivirus (HIV)
• Oncoviruses (Leukemia)
 Virus Families
9) Double-stranded RNA, Nonenveloped

• Reoviridae
• Reovirus (Also known as Respiratory Enteric
Orphan Virus)
• Rotavirus (Mild respiratory infections and
gastroenteritis)
• Colorado tick fever
 Growing Viruses
Plaque method
•Viruses must be grown in living cells.
•They are never grown in culture media
• Bacteriophages form plaques on a lawn of bacteria.
• Each plaque corresponds to a single virus in initial
suspension
• The No. of plaques are expressed in
Plaque-forming units (PFU)

Plaques onE. coli

 Growing Viruses
Animal viruses may be grown in
living animals or in embryonated
eggs.
• Vaccine virus in eggs

•Viral growth is signaled by the

death of the embryo, by embryo
damage, or by formation of
typical pocks or lesions on the
egg membranes
•Viral vaccine and egg allergy
(egg proteins)

Figure 13.7
• 1. Primary Cell Lines
Viral cultures
• From tissue slices
• die out after a few generations
2. Diploid Cell Lines
•
• derived from human embryos
• maintained for up to 100 generations
• Human diploid culture vaccine(rabies virus)
• 3. Continuous Cell Lines (immortal cell lines)
• Transformed Cells (Cancerous Cells)
• may be maintained indefinitely
• HeLa Cells
• Henrietta Lacks 1951 (Cervical Cancer)
• Propagation of virus not interfered with the changes in original cell
characteristics over years22
 Growing Viruses
• Animal and plants viruses may be grown in cell
culture.
• Continuous cell lines may be maintained
indefinitely.

Figure 13.8
 Virus Identification
Cytopathic effects CPE –

Negri bodies rabies

 Virus Identification
Serological tests
• Detect antibodies against viruses in a patient

• Use antibodies to identify viruses in neutralization tests, viral

hemagglutination, and Western blot
 Virus Identification
Nucleic acids
1) RFLPs (restriction fragment length polymorphism)

Restriction fragment length polymorphisms, or RFLPs, are differences among

individuals in the lengths of DNA fragments cut by enzymes. Restriction
enzymes are proteins that cut DNA at short, specific sequences called restriction
sites
 Virus Identification
Nucleic acids
2) PCR (polymerase chain reaction)
 Viral Multiplication
• Viruses don’t contain enzymes for energy
production or protein synthesis
• For a virus to multiply, it must invade a host cell and
direct the host’s metabolic machinery to produce viral
enzymes
• Multiplication of viruses can be demonstrated with a
one-step growth curve.
One-step Growth Curve
 Lytic cycle versus Lysogenic
The means by which a virus enters the host cell may vary but the
viral multiplication is similar for all viruses
Bacteriophages can multiply by two mechanisms:

•Lytic cycle Phage causes lysis and death of host cell

cell

•Lysogenic cycle Prophage DNA incorporated in host DNA

Multiplication of Bacteriophages
(Lytic Cycle)
 The length of DNA of phage is only about 6% of that of E. Coli but has over
100 genes in it
 APBMR – 5 distinct stages of Bacteriophage Lytic multiplication cycle
 Attachment - Phage attaches by tail fibers to host cell
 Penetration - Phage lysozyme opens cell wall, tail sheath contracts to force
tail, core and DNA into cell
 Biosynthesis - Production of phage DNA and proteins
 Maturation - Assembly of phage particles
 Release - Phage lysozyme breaks cell wall
 Bacterial Bacterial Capsid DN
cell wall chromosome A
Capsid

Sheath
Tail fiber
1 Attachment: Tai
Base plate
Phage attaches l
to host cell. Pin
Cell wall
Plasma membrane

2 Penetration:
Phage penetrates
host cell and injects
its DNA. Sheath contracted

Tail core

3 Biosynthesis: DNA is
copied and
capsomeres are
produced

Figure 13.10.1
 Tail
DNA

4 Maturation:
Viral components are
assembled into Capsid
virions.

5 Release:
Host cell lyses and
new virions are Tail fibers
released.

Figure 13.10.2
The Lysogenic Cycle

Figure 13.12
 The Lysogenic Cycle
Lysogeny results in
•Lysogenic cells are immune to reinfection by the same phage
• Phase conversion i.e. host may exhibit new properties (eg
Cornybacterium diptheriae)
• It makes specialized transduction possible
Generalized transduction: bacterial genes can be picked up in a
phage coat and transferred to another bacterium
Specialized transduction: only certain bacterial genes can be
transferred (gal genes)
 Specialized Transduction
gal gene
Prophage Bacterial DNA

1 Prophage exists in galactose-using host

(containing the gal gene).
Galactose-positive
donor cell gal gene
2 Phage genome excises, carrying with it
the adjacent gal gene from the host.

gal gene
3 Phage matures and cell lyses, releasing
phage carrying gal gene.

4 Phage infects a cell that cannot utilize

galactose (lacking gal gene).
Galactose-negative
recipient cell
5 Along with the prophage, the bacterial gal gene
becomes integrated into the new host’s DNA.

6 Lysogenic cell can now metabolize galactose.

Galactose-positive recombinant cell

Figure 13.13
Multiplication of Animal
viruses
Same as bacteriophage except uncoating

• Attachment - Viruses attaches to cell membrane

• Penetration - By endocytosis or fusion
• Uncoating - By viral or host enzymes
• Biosynthesis - Production of nucleic acid and proteins
• Maturation - Nucleic acid and capsid proteins assemble
• Release - By budding (enveloped viruses) or rupture
Attachment, Penetration, and
Uncoating

Multiplication of Animal Viruses

Multiplication (Biosynthesis) of DNA Virus
Papovavirus 1 Virion attaches to host cell
7 Virions are released
Host cell
DN
A Capsid

DNA 2 Virion penetrates

cell and its DNA is
6 Virions mature Cytoplasm
uncoated
Capsid enters
into nucleus

Capsid proteins

mRNA

5 Late translation;
capsid proteins
are synthesized

3 Early transcription and

4 Late transcription; DNA translation; enzymes are
is replicated synthesized
Pathways of multiplication used by various RNA
containing viruses
Pathways of Multiplication(biosynthesis) for RNA
Viruses
Biosynthesis of RNA viruses that use DNA
Maturation and Release of an enveloped virus by
budding

Figure 13.20
 Cancer
• Activated oncogenes transform normal cells into
cancerous cells.
• Viruses capable of inducing tumors in animals are called
oncogenic viruses or oncoviruses.
• The genetic material of oncogenic viruses becomes
integrated into the host cell's DNA.
• Transformed cells have increased growth, loss of contact
inhibition, tumor specific transplantation antigen(TSTA)
and T antigens in their nucleus.
 Oncogenic Viruses
 Oncogenic DNA Viruses  Oncogenic RNA viruses
 Adenoviridae  Retroviridae
 Heresviridae Viral RNA is transcribed to
 Poxviridae DNA which can integrate
into host DNA
 Papovaviridae
HTLV 1
 Hepadnaviridae •
HTLV 2
 Hepatitis B Virus(HBV) •
• Latent Viral Infections
• Virus remains in asymptomatic host cell for long
periods
• Cold sores(fever blisters) ..Simplexvirus
• Chicken pox(shingles) …Varicellovirus
• Persistent(chronic) Viral Infections
• Disease processes occurs over a long period,
generally fatal
• Subacute sclerosing panencephalitis(SSPE)
(measles virus)