Inoculation viruses into embryonated
chicken egg
Good pasture in 1931 first used the embryonated hen’s egg for
the cultivation of virus.
The process of cultivation of viruses in embryonated eggs
depends on the type of egg which is used.
Viruses are inoculated into chick embryo of 7-12 days old.
The egg used for cultivation must be sterile and the shell
should be intact and healthy.
For inoculation, eggs are first prepared for cultivation, the shell
surface is first disinfected with iodine and penetrated with a
small sterile drill.
After inoculation, the opening is sealed with gelatin or paraffin
and incubated at 36°c for 2-3 days.
After incubation, the egg is broken and virus is isolated from
tissue of egg.
Viral growth and multiplication in the egg embryo is indicated
by the death of the embryo, by embryo cell damage, or by the
formation of typical pocks or lesions on the egg membranes
Viruses can be cultivated in various parts of egg like
chorioallantoic membrane, allantoic cavity, amniotic sac and
yolk sac.
inoculation in embryonated eggs are:
1. Chorioallantoic membrane(CAM)
2. Amniotic Cavity
3. Allantoic Cavity
4. Yolk sac
1. Chorioallantoic Membrane (CAM):
Inoculation is mainly in 10-13 day
Inoculation is mainly for growing poxvirus.
After incubation and incubation, visible lesions called pocks are
observed, which is grey white area in transparent CAM.
Herpes simplex virus is also grown.
Single virus gives single pocks
This method is suitable for plaque studies.
2. Allantoic cavity:
Inoculation is mainly in 9-11 day
Inoculation is mainly done for production of vaccine of
influenza virus, yellow fever, rabies.
Most of avian viruses can be isolated using this method.
3. Amniotic sac:
Inoculation is mainly in 10-12 day
Inoculation is mainly done for primary isolation of influenza
virus and the mumps virus.
Growth and replication of virus in egg embryo can be detected
by haemagglutination assay.
4. Yolk sac inoculation:
Inoculation is mainly in 7 day
It is also a simplest method for growth and multiplication of
virus.
It is inoculated for cultivation of some viruses and some
bacteria (Chlamydia, Rickettsiae)
Immune interference mechanism can be detected in most of
avian viruses.
Advantages of Inoculation into embryonated egg
1. Widely used method for the isolation of virus and growth.
2. Ideal substrate for the viral growth and replication.
3. Isolation and cultivation of many avian and few mammalian
viruses.
4. Cost effective and maintenance is much easier.
5. Less labor is needed.
6. The embryonated eggs are readily available.
7. Sterile and wide range of tissues and fluids
8. They are free from contaminating bacteria and many latent
viruses.
9. Specific and non specific factors of defense are not involved in
embryonated eggs.
10. Widely used method to grow virus for some vaccine
production.
Disadvantages of Inoculation into embryonated egg
1. The site of inoculation for varies with different virus. That is,
each virus have different sites for their growth and replication.
