Hello, I want to do the infectivity assay to test SIV virus stocks using TZM-bl cell and luciferase assay. Is there a direct way to have TCID50 directly form RLU? Once I'm gonna have the RLU how do I calculate the TCID50? I know that there's an excel sheet out there but I can't find it!
Thanks for the help!
It's not as simple as plugging in your RLU result into a formula.
Think about the luminescence signal -- where does it come from? It comes from expression of a luciferase reporter gene in your TZM-bl target cells. The actual magnitude of the luminescence signal therefore depends on many factors: not just the number of infected cells in your well, but also the magnitude of luciferase expression, the specific luciferase assay reagents used, the sensitivity of your plate reader, etc.
TCID50, on the other hand, is an endpoint measurement to measure viral titer by asking how much virus is needed to cause infection of 50% of wells, and each well is scored only yes/no for infection.
Now, if you were to generate a standard curve of RLU vs pfu/mL or TCID50/mL using your own reagents, plate reader, etc, it might be possible then to convert RLU to TCID50/mL, but this conversion would apply only to a very specific set of parameters unique to your cells, reagents, plate reader, assay protocol, etc.
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