faster way to identify RNA virus other than RT-PCR? - (May/12/2006 )
hi, anyone knows if it is possible to identify a RNA virus through other methods other than reverse-transcription PCR which is not only relaible but also a quicker method??
and also does a band matching the size of a positive control in agarose gel electrophoresis tell us if the organism is dead or alive??
First question according to me depends on your virus. Certain virusses have a DNA intermediate, so you could isolate DNA (from cells) and amplify this (regular PCR). Others don't, but probably you could work with some Elisa technique (on a protein of your virus or an antigen in an infected individual).
Other than that there's NASBA and branched DNA techniques that are possible ass well, but it depends on what you want/need to do... If it's for diagnosis, you need a VERY consistent protocol with very good results, specificiy/sensitivity/reliability and so on... (definately with RNA virusses this is a problem because the lack of proofreading in their own polymerases).
The band matching the one in a positive control tells you nothing but having contamination OR indeed having virus, active or not cannot be told, but if your sample has inactive virus, you probably have active as well, not?
If this is something you'll do a lot of, maybe you can raise antibody to the virus, and use that to detect it in ELISA or western blot?