I am working with chicken anemia virus now, and I am using MSB1 cells. But it doesn't show clear and significant CPE, compare to negative control. So after I read some references, I did PCR for every dilution of the titration and read the result by gel electrophoresis. But then how to determine the TCID50 from the PCR result?
In PCR you see also destroyed virus particle, because the DNA is still there.
you see CPE only in wells with functional viral particle that can destropy the cells.
It is not only about the number of virus particle, but the number of "infectious particles"
If your TCID50 shows no significant CPE try a larger variety of dilutions in the range you expect the TCID50.
Good Luck
I agree with Ninon, PCR does not provide you the functional titer. Increase your particle number in the dilutions to see, if you can observe any CPE.
Hi,
Could you provide more information on how you infect CAV with MSB1 cells?
In addition to the informations mentioned above by Ninon
You advance in the passages of isolation until you will have a general CPE in the 24h of inoculation, like that you will have a great chance to have a high title therefore a good TCID50
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