INTRODUCTION TO VIROLOGY 2.

Icosahedral form
– Fixed or compact structure
Viruses
– Sturdy, hollow-core inside
 Smallest infectious agents (about 20nm to about
– 20 faces, each an equilateral triangle; 12 vertices,
300 nm in diameter)
– Capsids with icosahedral symmetry consist of a shell
 they depend on cells for molecular building blocks,
built from protein molecules that appear to have
machinery and energy
been arranged on scaffolding in the form of an
 infect all cellular life forms: eukaryotes (vertebrate,
icosahedron
animals, invertebrate animals, plants, fungi) and
– They have less contact with the virus genome
prokaryotes (bacteria and archaea)
3. Complex form
 Contain only one kind of nucleic acid as their
– Some virus particles do not exhibit simple cubic or
genome
helical symmetry , but are more complicated in
Structure:
structure
Virions
Type of nucleic acid
 viruses survive as virus particles outside their host – DNA or RNA, ss or ds, linear or circular
cells. – Usually in one piece, sometimes segmented
 gene delivery system; it contains the virus genome DNA Viruses
 Its functions are to protect the genome and to aid • Herspesviridae
its entry into a host cell, where it can be replicated • Hepadnaviridae
and packaged into new virions. • Adenoviridae
 The genome is packaged in a protein structure • Parvoviridae
known as a capsid • Papovaviridae
Capsid: • Poxviridae
 The protein shell, or coat, that encloses the nucleic Properties of DNA viruses
acid genome a. Resembles host DNA for transcription&
Functions of the capsid replication
 protection of the genome. b. Not transient nor labile
 to recognize and attach to a host cell in which the c. Viral genomes remain in the infected cells
virus can be replicated. d. Established persistent infection
 Stimulates the immune system to produce e. DNA genomes reside in the nucleus
antobody RNA viruses
Spikes or peplomers – Bunyaviridae
 Associated glycoprotein to an envelope – Arenaviridae
Envelope – Togaviridae
 Not produced by the virus – Orthomyxoviridae
 Made up of glycoprotein – Flaviviridae
 Sequestered by the virus upon exit – Filoviridae
– Paramyxoviridae
– Bunyaviridae
– Arenaviridae
– Togaviridae
– Orthomyxoviridae
– Flaviviridae
– Filoviridae
– Paramyxoviridae
– Picornaviridae
– Reoviridae
Classification : – Retroviridae
– Rhabdoviridae
 Type of capsid symmetry
– Caliciviridae
 Type of nucleic acid
– Coronaviridae
 Presence or absence of envelope
Characteristics of RNA viruses
 Polarity of the viral genome
– Genome structure determines the mechanism
International Committee on Taxonomy of Viruses
of transcription and replication
(2000)
– Labile and transient
 had organized more than 4000 animal and plant
– Replicate in the cytoplasm (mostly)
viruses into 56 families, 9 subfamilies, and 233
– Cannot replicate RNA
genera, with hundreds of viruses still unassigned.
– Prone to mutation
 24 families contain viruses that infect humans and
RNA viruses
animals.
– Majority are linear and majority are single-
Types of capsid symmetry
stranded
1. Helical form
– Circular (Arenaviridae)
– Seen in RNA viruses
– Families with segmented genomes
– RNA is coiled in the form of a helix and many copies
– Ortho- 7-8
of the same protein species are arranged around
– Rheo- 10-11
the coil
– Arena -2
– Provides flexibility in terms of shape of the capsid
– Bunya- 3
– Ability of the viral agent to undergo pleomorphism
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 Outer covering
Naked virus
Associated with acute infection
– Not easily damaged by adverse condition
– Can spread easily
– Do not cause persistent productive infection
– Exit by killing the host cell
Enveloped
– Less stable
– Must stay wet
– Cannot survive the GIT
– Causes viruses to be sensitive to acid and lipd
solvents
– Allows cell to cell fusion
Polarity of the viral genome
– In relation with mRNA
– Positive sense (+ sense): the same as mRNA
– Negative sense (- sense): compliment of mRNA
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Steps in Viral Replication
Adsorption- initial attachment of a virus to a host
 Temperature and energy dependent
 Reversible: if penetration does not ensue, the virus
can elute form the cell surface
 Irreversible: can lead to changes in virion structure
Penetration (endocytosis)
 Virus is engulfed by cell through invagination of
the cell membrane
 pH and temperature dependent
Penetration (fusion)
 Viral envelope merges with the membrane of the
host cell
 pH independent, temperature independent
Uncoating
 Stepwise process of the virion disassembly for the
purpose of releasing the viral genome
Replication
Assembly
Release
Lysis:
– For naked viruses
– Viral encoded toxic products program the death
of cell
Budding
– For enveloped viruses
– Cell not damaged
Methods Used In Virology
Specimen selection & collection
 Specimens for the detection of virus should be
collected as early as possible following onset of
symptomatic disease
 Virus may no longer be present as early as 2 days
after the appearance of symptoms
 Throat, nasopharyngeal swab or aspirate
 Throat- enteroviruses, adenoviruses, HSV
 Nasopharyngeal- RSV, influenza and parainfluenza
 Bronchial and Bronchoalveolar washes
– For viruses that infect the lower respiratory
tract (influenza and adenovirus)
 Rectal swabs and stool
– Rotavirus, enteric adenovirus and enteroviruses
 Urine
– CMV, mumps, rubella, measles, polyomavirus
and adenovirus
 Skin and Mucous membrane lesions
– Enteroviruses, HSV, VZV and rarely CMV or
poxviruses
 Sterile body fluids other than blood
– Enteroviruses, HSV, VZV, influenza or CMV
 Blood
– CMV, HSV and VSV
– 5-10ml in heparinized, citrated or EDTA for CMV
detection
– Citrated for other viruses
– Bone marrow
– Added to tube with heparin, citrate or EDTA
– Most viruses other than parvovirus B19 are
detected more readily from sites other than
bone marrow
 Tissue
– Viruses which infect the lungs (CMV, influenza
virus, adenovirus, sin nombre virus), the brain
(HSV), and GIT (CMV)
Specimen Transport and Storage
– All specimens collected must be processed
immediately
– Specimen for viral isolation must be placed in ice
and transported at once in the laboratory, if a delay
is unavoidable, the specimen shoulde be
refrigerated, NOT frozen until processing occurs
– Should be processes within 12-24 hours of
collection
– If specimen should be held for days before
processing
– 5 days: 40C
– 6 or more days: -20 or preferrably -700C
Blood for viral culture:
 Transported in a sterile tube with anticoagulant,
must be kept at refrigeration until processing
Blood for viral serology
 Transported in a sterile tube in which it was
collected
 Serum must be stored for hours or days at 40C or
for weeks or months at -200C or below before
testing
Cultivation
Animal cell culture
o well developed and most of the cells used are
from continuous cell
o lines derived from humans and other animal
species
o Continuous cell lines consist of cells that have
been immortalized, either in the laboratory or
in the body
Structural investigations of cells and virions
Light microscopy
 has useful applications in detecting virus-infected
cells, for example by observing cytopathic effect or
by detecting a fluorescent dye linked to antibody
molecules that have bound to a virus antigen
(fluorescence microscopy).
Electron microscopy
 The specimen, whether it is a suspension of virions
or an ultrathin section of a virus-infected cell, must
be treated so that details can be visualized.
 Negative staining techniques generate contrast by
using heavy-metal-containing compounds, such as
potassium phosphotungstate and ammonium
molybdate
X-ray crystallography
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 revealing detailed information about the three
dimensional structures of virions (and DNA,
proteins and DNA–protein complexes)
 Requires the production of a crystal of the virions or
molecules under study
Immunodiagnosis
 Virus antigens can be detected using virus-specific
antisera or monoclonal antibodies.
 Positive results are indicated by detecting the
presence of a label, which may be attached either
to the antivirus antibody (direct tests) or to a
second antibody (indirect tests)
Detection of virus nucleic acids
Hybridization
 Virus genomes or virus messenger RNAs (mRNAs)
may be detected using sequence-specific DNA
probes carrying appropriate labels
Polymerase Chain Reaction (PCR)
 When a sample is likely to contain a low number of
copies of a virus nucleic acid the probability of
detection can be increased by amplifying virus
 RNA can be copied to DNA and amplified using a RT
(reverse transcriptase)-PCR.
Virus genetics
Genome sequencing
 Determination of the sequence of bases in a
DNA molecule

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