Virus Detection - Tests for detecting specific virus (Feb/19/2007 )
I am working in research and development of viral vaccines.
Can some one please help me out in detecting the quantitative tests for detecting the virus after infecting it on to a cell base.
plaque assay, TCID50.
quantitative PCR? Alot more inaccurate, alot more difficult to start and a whole lot more expensive, but it can process larger volume of samples.
Hi avalon,
i am working on plaque assay with 2% agarose but i am not getting effective results. Can u suggest me some protocol.
Regards
Sandy
Hi Sandy,
What type of virus are you working with and what cell line? Are you getting any plaques? or CPE? I generally use 1 % agarose in my plaque assays. Have you tried checking for plaques ubder the microscope - they might be there but they may be very small. Ypu could try using tragacanth gum (final concentration of 0.75 %) instead of agarose. What are the other components of your overlay medium?
What type of virus are you working with and what cell line? Are you getting any plaques? or CPE? I generally use 1 % agarose in my plaque assays. Have you tried checking for plaques ubder the microscope - they might be there but they may be very small. Ypu could try using tragacanth gum (final concentration of 0.75 %) instead of agarose. What are the other components of your overlay medium?
Hi Avlon
I am working with MRC-5 and VERO cell lines. I can see the CPE as bulging of cells which is charecterstic of my virus.(measles). But when i go with Plaque assay i cant. at the same time can u suggest me weather i need to purify my virus before i go to any qualification or quantification tests such as Plaque assay, Heamagglutination.
Can some one please help me out in detecting the quantitative tests for detecting the virus after infecting it on to a cell base.
I am also working in viral vaccines. We use the Vero cell line too and use the plaque assay for quantification. Our first overlay is 1.8% agarose with the 2nd overlay being 1%