Diagnostic and treatment of Viral

infection
Muh. Nasrum Massi, MD., Ph.D
Makassar, Indonesia
Viral Diagnosis
 Diagnosis of viral disease
Specific clinical diagnosis
Supporting Laboratory:
1. Direct microscopic examination
2. Serologic examination
3. Viral isolation and identification
4. Experiments to animals
5. Molecular biology detection
 Specimen
Intact or new vesicle fluid
Swab/scraping from mouth ulcus,
from genitalia eye and bronchus eye
wash.
Saliva
LCS
Blood, feces, brain, back SST ,
abdominal organ
 Diluent for specimen
Nutrien broth
Sterile Aqua

Long distance shipment of specimen

uses glycerol 50% as preservative or
dry ice immediately into the
laboratory.
 Simple Test
With giemza staining or HE; basic
scratch of vesicle on base glass to
see giant cells with acidophilic
intranuclear inclusion body
With antibody fluorescence technique
inclusion body can be found quickly
inside the cell.
 Virus Isolation
1. In ovo inside embryonic egg
2. Invitro inside tissue culture cell
3. In vivo inside experimental animal
in the laboratory.
 In OVO
Better than in vivo because:
1. Egg buds cleanly and sterile, free of
bacteria
2. Does not have immunity mechanism like
experimental animal
3. Needless of food and cage
4. Chicken embryo have several amnionic
pouch, allantoic cavity, and corialantoic
membrane.
 Weakness of in OVO
Eggs could be contaminated by
mycoplasma & latent poultry virus
which could disturb the growth of
other viruses
Chicken embryo is sensitive only to
several kind of viruses
Just a subtle contamination to peem
substance could kill the embryo
 There are 3 kinds of tissue culture
(In vitro)
Culture of organs; culture of tracheal ring
used to isolate coronavirus
Explant culture, tissue slash is planted in
plasma clot. This technique is not applied
in virology anymore. Previously it was
used to culture adenoviruses.
Cell culture; most commonly used and
best technique, routinely applied in
virology
 Cell culture
Slices of cell tissue is released by
proteolytic enzym; tripsin ets or
mechanically shaken
Cells are washed, then suspended in
culture medium for culture then kept in
petri plates, reaction tubes, or bottles.
Cells adhere to the walls of the container
forming a monolayer and could be seen
with microscope with low magnification
 Laboratory results depend on:
1. Selection of the best specimen
(specimen of choice)
2. Collection, shipment, processing,
and storage of specimen
3. Selection and reduction of system
or live medium most sensitive for
the virus to be isolated.
 3 kinds of cell culture
1. Primary cell culture
Normal cells can be collected from the body
for the first time, can be transferred, and
cultured repeatedly.
Useful for viral isolation and culture for
making vaccines; kidney cell culture of
rhesus monkey, amnion cell culture of
human, chicken embryo fibroblast
culture, etc.
2. Diploid cell strain
Able to divide 100 times in culture.
Useful for isolation of pathogenic
virus which are difficult to culture
and also for vaccine production;
strains of human embryo lung cells
(WI-38) and strain of rhesus embryo
cells (HL-8)
3. Continous cell pathway
Group of cells that can divide unlimitedly as
One kind of cell is derived from cancer cells.
Could be cultured in several generation by
transferring from one tab to another. For
exp. HeLa cells (cell strain from human ca
cervix) KB (cell strain from human ca
nasoph) EP2 (cell strain from epitelioma)
Mc Coy (cell strain from human ca sinovia)
BAK21 (cell strain of hamster baby
kidney) and Detroid-6 (cell strain from
human chest SST)
 In vitro culture is useful for:
Primary virus isolation from clinical
specimens. Selected are cells with high
sensitivity, easy and fast to show ESP
Production of vaccine. Selected are cell
that produce a big amount of virus.
Biochemical investigation, selected are
usually continous cells in suspension
 Advantages and disadvantages of
rapid tests
Advantages
Provide quick results 30 minutes
Useful for detecting influenza outbreaks
May be used for clinical management
Simple procedures, variety of settings
Disadvantages
Less accurate than viral culture or PCR
Some tests do not distinguish A & B infection
Cannot identify influenza A subtypes
No data on strain, molecular or sequence
Test fail to pick up low titer H5N1 viruses
Interpretation of Rapid Test Results for
Suspected Avian Influenza A (H5N1)
Negative result does not = No H5N1
infection:
False negative (H5N1 detected by other
tests)
True negative (no influenza A virus
infection)
Positive result does not = H5N1 infection:
False Positive
Non-specific binding

Influenza B

True Positive (H1N1, H1N2, H3N2, H5N1)

 Summary of Serology

Detection of neutralizing antibody more sensitive

than detection of HI antibody against avian
influenza viruses in human sera
Micro-neut assay is valuable tool for:
outbreak investigations
Seroepidemiologic / surveillance studies

Need to address specificity issues

Detection of neutralizing antibody useful for

detection of vaccine responses to human
influenza viruses, e.g. vaccine strain selection
Treatment of Viral Diseases
 3 Methods
1. Chemotherapy
2. Immunization

3. Chemical which can produce

interferon or increase host defense
Another aspect:
1. Behavior

2. environmental
 Immunization

Too many vaccine have been

produced and effective in
implementation, such as; polio,
yellow fever.
Vaccine made by inactive virus,
genetic and synthetic.
Good vaccine if save in utilities and
effective
 In virus that cause respiratory tract
infection antigenic drift always occur and
the vaccine becomes ineffective
The majority of vaccine are derived from
live virus.
Vaccine could be gained naturally so tha
igG and IgA could be produced inspite of
local IgA
Vaccine could be tried on experimental
animal then on volunteers to see the
disappearance of viral virulence.
 Interferon
Antiviral substance released by
preinfective hospes cell and can be used
to prevent repeated infection in normal
animal cell
Main characteristic;
1. Extraordinarily active, usually combine
with other proteins in the serum.
Interferon is a protein kept at pH=2,
4oC, for 48 hours, will protect the cell
from viral infection.
2. Interferon is species specific, and formed
after viral or bacterial infection, or uptake
of particular chemicals.
3. Interferon does not inhibit viral
reporoduction but protects the cell from
the effect of the virus. Interferon
stimulate cell to from protein that inhibit
viral reproduction.
4. There are 3 kinds of human interferon; I
leucocyte from leucocyte, I fibroblast and
I immune from lymphocyte.
 Other drugs
Levamisol and isoprinosin In-osiplex)
As immunostimulant
Effective against RNA and DNA virus
Amonium 21 tungsto 9 antimonate
(HPA) suramin have been tried
against HIV AIDS
Certain staining reagents
Application of stain and illumination
to kill newly formed infective virus