Hey Rick,
I think PCR is almost certainly the way to go, for a few reasons.
Firstly, you're talking about mammalian viruses; to grow them (which is
a bad idea in any case), you'd need mammalian cells, and mammalian cells
call for more intensive care than bacteria. It's a big investment, and
it requires a lot of time and effort just to keep them going.
And that's ignoring the fact that each virus will call for a different
target cell type, which in turn calls for different care procedures;
kidney cells may need very different medium and care than liver cells,
for example.
Secondly, plaque assays don't really work, AFAIK, for mammalian viruses.
Because they must be grown with a liquid layer on top, and that liquid
layer allows viruses to spread freely, instead of growing outwards like
bacteriophages.
Besides, if dealing with human infections, a good rule #1 is *never
culture*. If you can do your work without culturing the viruses at all,
then you should.
DNA survives boiling and alcohol, but viruses don't. If you want to
study your kids' colds, you could just take a sample of nasal mucous,
boil it in an eppendorf tube for 10 minutes, add 40% EtOH just to be
sure and then use a tiny sample for PCR, using primers that will amplify
your virus of interest. Ideally use primers that are already tested and
shown to work reliably in the available science literature. If you make
your own, aim for 25 nucleotides in length in a non-repetitive area of
the virus genome that's probably well-conserved, such as promoters.
PCR is great for many reasons. One reason is that it allows you to study
something that is impossible or unwise to culture, such as thermophiles,
bizzarre soil symbiotes, or human pathogens. I would strongly suggest
sticking with PCR and taking normal precautions to avoid infecting
yourself while you take samples.
On 25/01/12 05:51, Rick Byers wrote:
> Hi,
> I'm curious to better understand the pattern of viral infections in my
> family (I've got two young kids, so it's pretty common for some cold
> to be going around our house). I'm trying to figure out the most
> practical way to do regular URT diagnostics. I've got no lab
> experience (but would love to learn) and almost no lab supplies (but
> am willing to invest up to a few thousand dollars as necessary).
> Ideally I'd like something that is both quantitative (so I can track
> viral load over the course of infection) and multiplexed (so I can
> identify which viruses are responsible and catch most common ones).
>
> I know I can do some simple diagnostics at home with a modest
> investment. Eg., I considered doing simple plaque assays or simple
> PCR runs, but I'm hoping I can find something that's less time
> consuming at non-trivial scale (eg. detecting at least 10 different
> viruses/strains, ideally many samples over a period of time). Perhaps
> there's an immunoassay panel of some sort I could use at home? Or
> perhaps the best option is a service that will do a multiplexed qPCR
> run for me on a set of samples I send in.
>
> I'm continuing to do research of various products and diagnostics
> services, but I'm finding it pretty difficult to get specific prices
> and weigh the options. Any advice would be greatly appreciated.
> Thanks!
>
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