How to detect virus in organ? - (Jul/08/2008 )
Dear collegue scientist,
I would like to study a virus shedding in mice organs for the safety evaluation of the virus that purposely injected as a viro-onco therapy.
Can any body suggest the method especially for brain?
We usually cull the mice, remove the organs, homogenise and then detect virus by PCR and plaque assay.
Alternatively you could try in-situ pcr, but I have no experience with this so can't tell you how to go about that!
Thank you for the suggestion, do you have a refference book to suggest for the homogenise and isolation? TQ
For homogenizing you can use a homogenizer or use a cell strainer and the back of a plunger (the thing that's inside a syringe, which you use to draw the volume up).
Just mesh the tissue. Depends on which tissues you use and how many on what you prefer. In my old lab we never had a homogenizer and we used to do everything by hand, now I'm loving the homogenizer.!
Tissues like spleen and liver are the easiest. Lungs a little harder, mostly in the homogenizer, because they sometimes sit above the pedals and don't get homogenized. Use cell culture medium or you can use PBS probably if you're just looking at virus and not specifically at the cells.
Hope this helps a little.
[Edit] Check papers that check for virus titers in tissues. There lots of them out there.
Thank you and i found a lot of option for virus titer in tissue evaluation. But if also want to evaluate the virus presenting on paraffin embeded organ tissue, is it any suitable fluorescence method of detection?