After you isolated what seems to be an intracellular parasit

After you isolated what seems to be an intracellular parasite from a blood culture from a human case of chronic fatigue syndrome. You do an Electron microscopy, which indicates that the isolate looks like a virus, but the results are not conclusive. You now believe that this isolate is either a new virus or a protozoan parasite. Your virology professor tells you that your isolate is most likely a virus.

Explain in detail.

How can use eggs to grow this pathogen in large quantities and purify it? What would you have to do?

How would you go about using eggs to establish that the isolate is a virus?

How would you determine that you do have a virus?

Give detailed explanation and reasoning.

Solution

Using eggs to grow virus in large quantities:

1. The egg used for cultivation must be sterile and the shell should be intact and healthy.

2. 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.

3.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.

Purification of virus:

Viruses can be purified in various parts of egg like chorioallantoic membrane, allantoic cavity, amniotic sac and yolk sac.

Confirmation whether prtozoa or virus:

Firstly,recognized by EM, confirmed by other techniques, including serologic testing, immunohistochemical (IHC) and indirect fluorescence antibody (IFA) assays, and molecular methods that can further characterize the virus to species and strain.