Hi everyone,
When we amplify viruses in cell culture, definitely, all viruses produced can not be able to infect the new cell. There are always some failed events in viral production, which remain in the media without causing any further infection. Actually, I want to determine a ratio between live and dead viruses, in one of my experiments. So, possibly how can I do that.
Regards
Do you mean gene therapy-type viruses, or live infectious non-recombinant virus? If your question is regarding lentivirus, then as far as I'm aware it's not possible to actually separate out non-functioning virus. However, you can use an ELISA to look for all virus particles and then use either QPCR or microscopy to get an infectious titre - so you can get a ratio from there.
Hi there,
You may want to have a look at this: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4139191/
While mostly on ways to quantify virus on a particle basis, we also discuss the conundrum of infectious to total virus counts...
Hope that helps a bit,
CHristoph
Thank you Jill and Christoph for your response,
Jill, actually I am concerned with the wild type infectious viruses... Yup, we can use ELIZA. QPCR and microscopy... but, I have figured it out that I can also do with qPCR and Plaque assay in my case... Plaque Assay will give the viable count of the viruses while qPCR is good to determine total amount of viruses...
Christoph, I have started reading your paper... It will be truly very helpful for me in my research...
Regards...
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