Introduction to

Virology

AhmedSayedAbdelMoneim
Professor,MicrobiologyDepartment(Virology)
CollegeofMedicineandMedicinalSciences
TaifUniversity SaudiArabia

Selected Reading
Microbiology (Richard A.
A Harvey,Pamela
Harvey Pamela C
C. Champe)
Medical Microbiology (Introduction to infectious
diseases, Sherris)

Fundamental Virology (Fields)

Prof. Ahmed Sayed Abdel-Moneim

Objectives
To know the definition of the virus
To know the components
p
and the ggeneral functions of each
virus structure
Classification and nomenclature of viruses
To know how virus replicates inside the host cell
Types of virus infections and how virus infection could be
prevented
Diagnosis of virus infection

Prof. Ahmed Sayed Abdel-Moneim

Viruses
Viruses are obligate intracelluar parasites that are
metabolically
t b li ll inert
i t outside
t id their
th i hosts.
h t
They can multiply only in living cells.

Prof. Ahmed Sayed Abdel-Moneim

Virus: Structural components

Nucleocapsid
1.Nucleic acid
2.Protein coat [capsid]
3.Envelope in some viruses only

Prof. Ahmed Sayed Abdel-Moneim

Nucleic Acid

Viral Genomes
DNA

Single Stranded

Parvovirus

Double Stranded

Herpesviruses

Partial Double Stranded

Hepatitis B virus

Double Stranded

RNA
g Stranded
Single

Linear

Segmented

Positive sense

Poliovirus

Negative sense

Measles

Influenzavirues (Negative sense)

Prof. Ahmed Sayed Abdel-Moneim

Viral Capsid
It coats the viral nucleic acid
Significance of capsid?
Protect nucleic acid
Attach to receptors on cells
Capsid proteins provides antigen to which the
immune system react

Prof. Ahmed Sayed Abdel-Moneim

Types of Capsid
Helical symmetry

Icosahedral symmetry

Mixed

Influnezaviruses

Polioviruses

Poxviruses

Prof. Ahmed Sayed Abdel-Moneim

Viral Envelope
Some viruses ppossess envelope.
p
The envelope is cell integral part.
It consists of lipid bilayer [cellular origin ] studded by
viral epitopes [viral origin].
The envelope render viruses sensitive to lipid solvents.
Examples of enveloped viruses
Non enveloped viruses are called naked ones.

envelope

Prof. Ahmed Sayed Abdel-Moneim

Virus multiplication (replication)

It includes the following:

(1) Virus Attachment or adsorption
(3) Virus multiplication
(5) Virus Release

(2) Virus penetration and uncoating

(4) Virus Assembly

All DNA viruses replicate in the nucleus of the host cell except poxvirused replicate in
cytoplasm
All RNA viruses replicate in the cytoplasm except orthomyxoviruses and retroviruses
replicate in the nucleus.

Prof. Ahmed Sayed Abdel-Moneim

A) Virus attachment or adsorption

((a)) C
Collision.
lli i
R d
Random
movementt andd meeting
ti off virus
i particles
ti l andd host
h t cell
ll occurs
(b) Ionic attraction. Counter positive ions are needed.
(c) Receptor Binding:
Adsorption of the virus occurs to specific receptor sites on the surface of the susceptible
host cell is called (tropism).

Prof. Ahmed Sayed Abdel-Moneim

B) Penetration and uncoating

a-Penetration of necked viruses:
Endocytosis or phagocytosis. Through the endosome or the phaghocytic
vacuole.
Translocation. Viruses pass directly to inside the host cell without noticeable
changes
g in cell membrane as ppicorna viruses.
b-Penetration of enveloped viruses:
Endocytosis or phagocytosis.
Fusion to plasma membrane. Virons bind to cell surface receptors via a viral
epitope (surface protein).

Uncoating.
It means liberation of viral genome inside the host cell with subsequent
expression of viral coding genes.
Eclipse phase It is the phase following uncoating stage of viruses during which
viruses are not found inside the cell.

Prof. Ahmed Sayed Abdel-Moneim

Prof. Ahmed Sayed Abdel-Moneim

Prof. Ahmed Sayed Abdel-Moneim

Prof. Ahmed Sayed Abdel-Moneim

C) Virus multiplication
RNA viruses
All RNA viruses replicate inside the cytoplasm except retroviruses.
Single stranded RNA (+)sense non segmented
The genome acts as m RNA and translated directly in ribosomes.
Single stranded RNA (-)sense non segmented
p
RNA ppolymerase
y
(transcriptase)
(
p
) by
y which complementary
p
y positive
p
strand ((mRNA ) is
It carries RNA dependent
transcribed
Single stranded RNA (-)sense segmented
It carries RNA dependent RNA polymerase (transcriptase) by which complementary positive strand (mRNA ) is
transcribed for each segment
Single stranded RNA (+) sense segmented
Ex. retroviruses
reverse transcriptase: RNA to dsDNA
The resulted ds DNA is then integrate itself into host cell genome.
Segmented dsRNA
Ex. reovirus and birnaviruses
The genome is transcribed by virus associated transcriptase enzyme to give mRNA from the ve
ve strand
strand.
The mRNA molecules

Prof. Ahmed Sayed Abdel-Moneim

DNA viruses
All DNA viruses replicate inside the DNA except poxviruses
Parvovirus and hepadenavirus use reverse transcriptase enzyme

Prof. Ahmed Sayed Abdel-Moneim

Prof. Ahmed Sayed Abdel-Moneim

Prof. Ahmed Sayed Abdel-Moneim

D) Virus assembly
Capsid proteins are synthesized by host cell ribosomes
Empty procapsids are produced
Nucleic acid is inserted
Enveloped virus nucleocapsids are assembled similarly
E) Virus Release
Naked viruses are usually released when host cell lyses
Enveloped viruses are usually released by budding:

Prof. Ahmed Sayed Abdel-Moneim

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Consequences of infection
Productive: Production of progeny virus particles.
Abortive: Failure of infection
Latent:
The viral nucleic acid persists in a dormant state in certain cells after the virus
shedding. e.g. herpes viruses. The virus is reactivated when the host is
stressed.

Transforming:
Transform normal cells to tumor ones e.g. Retroviruses

Persistent
In this form, the virus is continuously detected with mild or no clinical
symptoms, e.g. chronic hepatitis B.
Prof. Ahmed Sayed Abdel-Moneim

Antiviral Therapy
I. Antiviral Drugs (modes of action):

Inhibition of uncoating
Inhibition of viral RNA polymerase
Inhibition of viral DNA polymerase
Reverse transcriptase inhibitors
Protease inhibitors
Inhibition of virus release

Interferons (IFNs, type I)

Prof. Ahmed Sayed Abdel-Moneim

11

I.

Antiviral Drugs

1- Inhibition of Uncoating
Amantadine/rimantadine: Inhibits uncoating of influenza A virus,
therefore used in prevention of its infection.

Prof. Ahmed Sayed Abdel-Moneim

Antiviral Drugs
2- Inhibit Viral DNA Polymerase
1- Acyclovir
y
((Zovirax):
)
e.g. active against HSV-I, II and Varicella Zoster virus.

- Topical acyclovir.
-Parenteral acyclovir.
- Oral
2- Ganciclovir:
similar
Cytomegalovirus [CMV].

to

acyclovir

but

more

effective

against

3 Inhibit RNA polymerase

3 Ribavirin: It inhibits synthesis of mRNA.

Prof. Ahmed Sayed Abdel-Moneim

12

Antiviral Drugs

4- Inhibit Reverse transcriptase [RT]

Zidovudine, AZT: It inhibits viral RT enzyme. It is used in treatment of
AIDS.
Lamivudine (3 TC) and Stavudine (d4T): Both inhibit RT enzyme of HIV

Prof. Ahmed Sayed Abdel-Moneim

5- Protease Inhibitors
Proteases are needed in the late stage of HIV replication.
Indinavir is a model of protease inhibitors.

6- Inhibition of Virus release: Tamiflu oseltamivir

Prof. Ahmed Sayed Abdel-Moneim

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II. Interferons (IFNs)

These are host coded proteins (cytokines) that inhibit viral
replication.
They are the first line of defense against viral infections.

Prof. Ahmed Sayed Abdel-Moneim

Diagnostic Methods in Virology

1. Direct Detection
2. Virus Amplification
3. Serology

Prof. Ahmed Sayed Abdel-Moneim

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Direct Detection
1. Virus particles By Electron Microscopy

Prof. Ahmed Sayed Abdel-Moneim

2. Detection of Inclusion bodies by Light Microscopy

Normal stains as Giemsa or H&E

Special stains as Sellers stain as in rabies virus

Prof. Ahmed Sayed Abdel-Moneim

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3. Antigen Detection
Cell associated Ag
immunofluorescence,
immunoperoxidase ,
cell ELISA

Soluble Ag ELISA, AGPT[agar gel

preciptation
i t ti test],
t t] etc
t

Prof. Ahmed Sayed Abdel-Moneim

4. Viral Genome Detection

Insitu nucleic acid detection In situ hybridization
Purification of nucleic acid and detection by
Dot blot hybridization
Restriction fragment length polymorphism analysis[RFLP]

Prof. Ahmed Sayed Abdel-Moneim

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Virus Amplification
A) In-vivo
In vivo
1. Cell Culture

2. Embryonated Eggs

3. Animals
Prof. Ahmed Sayed Abdel-Moneim

Types of Cell Culture

1. Primary cell lines:
2. Human diploid cell culture: Human embryonic lung
3. Continuous cell lines:
Prepared from
Tumor cells.

Prof. Ahmed Sayed Abdel-Moneim

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Detection of Virus Replication in Cell Culture

1. Cytopathic effect (CPE):
- Cell death or lysis.
- Syncytial formation (giant cells):
Fusion of membranes of adjacent cells to form multinucleated giant cells.
- Inclusion bodies:
These are either the site of virus assembly or degenerative changes in the cell.
a- Intracytoplasmic: e.g. rabies.
b- Intranuclear: e.g. herpes viruses.
c- Both: e.g. measles virus.

-Plaque
Pl
f
formation:
i
Plaques are areas of virally-infected cells in a cell culture.

Prof. Ahmed Sayed Abdel-Moneim

Viral Plaques

Prof. Ahmed Sayed Abdel-Moneim

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In vitro amplification
PCR

Prof. Ahmed Sayed Abdel-Moneim

Serology
Detection of rising titres of antibody between acute and
convalescent stages of infection, or the detection of IgM in
primary infection.
Classical Techniques

Newer Techniques

1.
2
2.
3.
4.
5.

1.
22.
3.
4.
5.

Complement fixation tests (CFT)

Haemagglutination inhibition tests
Immunofluorescence techniques (IF)
Neutralization tests
Counter-immunoelectrophoresis

Radioimmunoassay (RIA)
Enzyme linked immunosorbent assay (EIA)
Particle agglutination
Western Blot (WB)
RIBA, Line immunoassay

Prof. Ahmed Sayed Abdel-Moneim

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Serology
Criteria for diagnosing Primary Infection

4 fold or more increase in titre of IgG or total antibody

between acute and convalescent sera
Presence of IgM
Seroconversion
A single
i l high
hi h titre
i off IIgG
G ((or totall antibody)
ib d ) - very
unreliable

Prof. Ahmed Sayed Abdel-Moneim

Typical Serological Profile After Acute Infection

Note that during reinfection, IgM may be absent or present at a low level transiently
Prof. Ahmed Sayed Abdel-Moneim

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Complement Fixation Test

Complement Fixation Test in Microtiter Plate.

Prof. Ahmed Sayed Abdel-Moneim

ELISA for antibody

Microplate ELISA for HIV antibody: coloured wells indicate reactivity

Prof. Ahmed Sayed Abdel-Moneim

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