Method of cultivation of

:viruses
Since viruses are obligate
intracellular

parasites, they have to be grown in living cells.

: There are three systems for their cultivation

(1)

Laboratory
animals

(2)

Embryonated
eggs

(3)

Cell culture

Method of cultivation of
viruses
:Laboratory animals
Some animal viruses can be cultured only in
.living animals, such as mice, rabbits, and pigs
Most

experiments

to

study

the

immune

system's response to viral infections must also

. be performed in virally infected live animals
After

the

animal

is

inoculated

with

the

specimen, the animal is observed for signs of

disease or is killed so that infected tissues can

Method of cultivation of
Some human viruses
cannot be grown in
viruses
animals or can be grown but do not cause
.disease

Method of cultivation of
viruses
:Embrionated eggs.2
If the virus will grow in an embryonated egg,
this

can

be

fairly

convenient

and

inexpensive form of host for many animal

.viruses
A

hole

is

drilled

in

the

shell

of

the

embryonated egg, and a viral suspension or

suspected virus-containing tissue is injected
. into the fluid of the egg

This method was once the most widely used

method of viral isolation and growth, and it is
still used to grow viruses for some vaccines. For
this reason, you may be asked if you are allergic
to eggs before receiving a vaccination, because
egg proteins may be present in the viral vaccine
.preparations

Method of cultivation of
viruses
:In Cell Cultures .3
Cell cultures consist of cells grown in culture
.media in the laboratory

Cell culture lines are started by treating a

slice of animal tissue with enzymes that
.separate the individual cells
These cells are suspended in a solution that
provides the osmotic pressure, nutrients,
and growth factors needed for the cells to

Method of cultivation of
viruses
:In Cell Cultures .3
Normal cells tend to adhere to the glass or
plastic container and reproduce to form a
.monolayer
Viruses

infecting

such

monolayer

sometimes cause the cells of the monolayer

.to deteriorates they multiply
This cell deterioration is called cytopathic
, effect (CPE)

Diagnosis of virus
infections
Thelaboratory
laboratoryprocedures
procedures
usedin
in
The
used

:diagnosisof
ofviral
viraldiseases
diseasesinclude
include
:diagnosis

)A(

Direct detection
of viruses

)B(

Serologic
detection of
antiviral
antibodies

Direct detection of viruses(A)

Direct detection of viruses can be Achieved by
:different techniques
:Light microscopy .1
:Light microscopy .1
This can be used to
This can be used to
visualize some large
visualize some large
viruses e.g. poxviruses
viruses e.g. poxviruses

:Electron microscopy. 2
Is used to demonstrate virus particles in
vesicular fluid or tissue extracts treated with
special stain. Its only successful if large
.numbers of particles are present

:Immunofluorescence microscopy. 3
Detection of virus in smears from lesions
using fluorescein labeled specific antisera

:Polymerase chain reaction. 5

This technique involves amplification of a
short sequence of a target DNA or RNA (which
may be in low concentration e.g. one copy)
leading to accumulation of large amounts of
.that sequence, so it can be easily detected
PCR can be used to determine the quantity of
viruses in patients blood i.e. virus load e.g. in
HIV patients. This is used to monitor the
course of the disease and to evaluate
. treatment and prognosis

:Serologic detection of antiviral antibodies(B)

:Serologic diagnosis used in virology
:Enzyme linked immunosorbent assay(ELISA). 1
:Enzyme linked immunosorbent assay(ELISA). 1
Enzyme linked immunosorbent assay (ELISA)
Enzyme linked immunosorbent assay (ELISA)
can be used for detection of viral antigens in
can be used for detection of viral antigens in
different clinical specimens e.g. detection of
different clinical specimens e.g. detection of
.HBV ,HIV, HCV and Rota virus
.HBV ,HIV, HCV and Rota virus