Agent of infection:

The Viruses
Purnomo Hadi
Dept. of Microbiolgy FK UNDIP – RSUP dr. Kariadi
Introduction
 Are obligate intracellular parasites of bacteria, protozoa,
fungi, algae, plants, and animals.
 Ultramicroscopic size, ranging from 20 nm up to 450 nm
(diameter)
 Lack enzymes for most metabolic processes
 Lack machinery for synthesizing proteins
Comparative
sizes of virions,
their nucleic
acids, and
bacteria.
Acellular(virus) – Procaryote - Eucaryote
 VIRUSES’s STRUCTURE

Naked &
Enveloped Virus
VIRUS
Morphology
 Components of Viruses

1. Genome
 Nucleic acid
 can be either DNA or RNA but not both

2. Capsid
 protein coat is composed of a number of individual protein
molecules,
 arranged in a precise and highly repetitive pattern around the nucleic
acid
 constructed from identical subunits called capsomers
Components of Viruses

3. Envelope
 membrane surrounding the nucleocapsid
 consists of a lipid bilayer with proteins, usually glycoproteins, embedded in it
 derived from the membranes of the host cell

4. Spikes (Peplomer)
 Some proteins form a binding layer between the envelope and capsid
of the virus
 glycoproteins remain exposed on the outside of the envelope form
protruding molecules,
 essential for the attachment of viruses to the next host cell
Classification:
Classification:
 Genetic’s Dogma

DNA transkripsi RNA translasi Protein

Functional
replication

Reverse
transcriptase
DNA
Central dogma of molecular biology
 Unusual flow of information highlighted in green
 Baltimore Classification of Viruses, base on viral genom
and replication mode

Group Genome Replication Example

1 dsDNA dsDNA mRNA Herpes simplex
virus

2 ssDNA ssDNA dsDNA mRNA Parvovirus

3 dsRNA dsRNA mRNA Reovirus

4 +ve ssRNA dsRNA +ve ssRNA [Acts as mRNA] Enterovirus

5 -ve ssRNA dsRNA -ve ssRNA mRNA Influenza A

virus

6 ssRNA ssRNA dsDNA mRNA Retrovirus

(e.g. HIV)

7 Nicked dsDNA nicked dsDNA intact dsDNA mRNA Hepatitis B

virus
RNA
Virus Host
UNCONVENTIONAL AGENTS
('unconventional viruses' or 'atypical viruses„)
1. VIROIDS
 defective or satellite viruses: dependent on other viruses for
replication
 small (<400 nucleotides), single stranded, circular RNAs. Non-
envelopes
 Example: adeno-associated virus (AAV), delta agent (Hepatitis D virus)

2. PRIONS
 composed primarily of protein (no nucleic acid)
 smaller and simpler than viruses
 implicated in chronic, persistent diseases in humans and animals
 These diseases are called spongiform encephalopathy's. because the
brain tissue removed from affected animals resembles a sponge
 Examples: Creutzfeldt-Jakob disease (human), Bovine spongiform
encephalopathy/ “mad cow disease,”
Stages in Virus-Host Interaction
1. Entry into the host
2. Primary replication
3. Spread
4. Cell and tissue tropism
5. Secondary replication
6. Cell injury or persistence
7. Host immune response
Tissue Tropism
 Viral affinity for specific body tissues (tropism) is
determined by:
1. cell receptors for virus,
2. cell transcription factors that recognize viral promoters and
enhancer sequences,
3. ability of the cell to support virus replication,
4. physical barriers,
5. local temperature, pH, and oxygen tension enzymes and
non-specific factors in body secretions, and
6. digestive enzymes and bile in the gastrointestinal tract that
may inactivate some viruses
Viral receptor:
Viral entry: fussion
Viral entry: endoctytosis
Bakteriofag T4: Lytic Cycle
Bakteriofag T4: Lysogenic Cycle

Replication
Cellular Pathogenesis
 Direct cell damage and death from viral infection may
result from:
 diversion of the cell's energy,
 shutoff of cell macromolecular synthesis,
 competition of viral mRNA for cellular ribosomes,
 competition of viral promoters and transcriptional enhancers
for cellular transcriptional factors such as RNA polymerases,
and
 inhibition of the interferon defense mechanisms.
 Indirect cell damage can result from integration of the
viral genome, induction of mutations in the host genome,
inflammation, and the host immune response.
The pathologic effects of viral diseases
 toxic effect of viral genes products on the metabolism of
infected cells,
 reactions of the host to infected cells expressing virus
genes, and
 modifications of cellular functions by the interaction of
cellular DNA or proteins with viral gene products
Incubation Period
 is the time between exposure to virus and onset of
disease
 During this:
 asymptomatic period,
 implantation,
 local multiplication, and
 spread (for disseminated infections) occur.
Local Replication and Local Spread
 Most virus types spread among cells extracellularly, but some
may also spread intracellularly.

Dissemination from the Portal of Entry

 Viremic:
 The most common route of systemic spread from the portal of entry
is the circulation, which the virus reaches via the lymphatics.
 Virus may enter the target organs from the capillaries by:
1. multiplying in endothelial cells or fixed macrophages,
2. diffusing through gaps, and
3. being carried in a migrating leukocyte.
 Neural:
 rabies virus and
 sometimes with herpesvirus and poliovirus infections.
Virus local infection
Virus systemic infection: Viremia
Systemic infection: neural spreading
Multiplication in Target Organs
 Depending on the balance between virus and host
defenses, may be sufficient to cause:
 disease
 death
Shedding of Virus
 the most frequent sites of shedding:
 alimentary tract,
 urogenital tract and
 blood are
Congenital Infections
 the fetus as a target "organ" is special:
 virus must traverse an additional physical barriers (placenta),
 the developing first-trimester fetal organs are vulnerable to
infection, and
 hormonal changes are taking place.
 Example:
 Rubella
 CMV
Kinds of infection:
1. Infection:
1. Acute infection
2. Persistent Viral Infections:
a. latent,
b. Chronic
c. slow infection.

2. Oncogenic Transformation
3. Virus-Induced Immunopathology
Problems in Virology
 Identification and diagnostic of viral diseases:
 Intracellular obligate  difficulty to culture
 Therapy in viral diseases:
 Intracellular obligate  using almost all of host enzymes
 Control of viral diseases:
 Speed of transmission
 Asymptomatic diseases
 Unique pathogenesis
 Zoonosis
 Influenza
 Influenza pandemics, death case:
 H1N1 (1918): Spanyol  50 million?
 H2N2 (1957): Asian  1-2 million
 H3N2 (1968): Hongkong  1 million

39 Purnomo Hadi
Dengue
infection…..
DIAGNOSTIC STRATEGY

TBC TBC
HZV

Oral candidiasis
PCP OHL
Cryptococcus diarrhea Cryptococcus meningitis
PPE
CMV MAC
Rubeola/measless - Mumps
Rubeola/Measless - Rubella
Hepatitis
 Very high genetic variation

 6 genotypes (similarity within genotypes 91%)

 > 50 SUBTYPES (similarity within 1 genotype

similarity 77 – 80%)

 Similarity among genotypes (66-69%)

Varicella & Herpes zooster virus
Herpes simplex virus
Polio
 Papilloma: warts – Condyloma acuminata

Cervical cancer?
Papilloma: Dede, “The Roots Man”
Viral Identification
Algoritme of
Diagnosis of
Infetion
DIAGNOSIS of INFECTIONS

1. Detect agent of infection

1. Whole organism:
1. Microscopic examination
2. Culture
2. Parts of organism:
1. Antigen
2. DNA
2. Detect immune response of infection
 Light Microscopy
 Detect CPEs: morphology
changes, cell lysis, vacuolation,
synctia and inclusion bodies (IB).
 Cynctia are multinucleated giant
cells e.g. HSV, HIV and
Paramyxoviruses.
 IB histological changes in cells
induced by viral components:
cowdry A inclusion by HSV,
owl’s eye by CMV and negri
bodies by rabies
 Electron Microscopy

 Usual magnification : 50,000

 Mainly used for the diagnosis of viral gastroenteritis
 Expensive
 Poor sensitivity (106 virus particles/ml specimen)
 Need highly skilled observer
 Becoming less widely used due to availability of reliable
antigen detection and molecular methods

Norwalk-like
viruses
2. Virus Isolation /culture:

 Specimen - inoculated into cell culture, eggs or

animals
 Eggs and animals - difficult to handle
 most viral diagnostic laboratories - cell culture only.

3 types of cell cultures:

a. Primary cells
b. Semi-continuous cells
c. Continuous cells
Immuno-fluorescence assay:
Figure 3. Specimen of Lung Tissue from the Index Patient's Mother.
Immunohistochemical analysis of the specimen shows interstitial pneumonitis and a single
epithelial cell containing intranuclear influenza A viral antigens (red) and an
antinucleocapsid antibody.

Antigen detection:
Immunocyto-
chemistry
Antibodies Detection:
Immune response
Molecular: PCR
Thank you……..