Can anyone share TCID50 protocol for rubella virus titration using vero cells?

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Why my sandwich ELISA shows half the concentration of injected unknown?

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Is it a true TCID50 titration if I had freeze the plates after 11 days and ran ELISA later to titer Rubella antigen?

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Can anyone share TCID50 protocol for rubella virus titration using vero cells?

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Ahmed H. El-Sappah

Hello. I suggest you these protocols :-

1- TCID50 Assay Protocol

http://www.virapur.com/protocols/TCID50%20Protocol.pdf

2- Detection of measles, mumps and rubella viruses by immuno-colorimetric assay and its application in focus reduction neutralization tests

Article Detection of measles, mumps and rubella viruses by immuno-co...

3- VIROLOGY MANUAL OF THE DEPARTMENT OF NEUROVIROLOGY, NIMHANS, BANGALORE 560029 OF INFECTIONS OF THE NERVOUS SYSTEM

Aakash Patel

Thanks Ahmed H. El-Sappah for the reply. I was wondering if it would be possible to amplify the rubella virus in cell culture plate- infecting Vero cells in the plates and after 4 days of infection transfer the supernatant to another Vero cell seeded plate. Subsequently, after 4 days, collect supernatant and run ELISA.

Do you think this idea can work?

Karen A. Darbinyan

Hello, the reason for all this?

But if you want yes you can amplify the rubella virus in Vero cell culture as you mentioned above only thing 4 day may be more than required. If in 4 day you have max titer of virus so it better to transfer in to the new portion of cells the day earlier and than wait 4 days.

Good luck.

Thank you Karen A. Darbinyan. As you may know TCID50 in Vero cells are not most ideal method to determine Rubella virus titer so, we want to develop an assay that provides high-throughput.