An obscure intracellular agent etc...a prank??

Karl Fischer kfischer at gpu.srv.ualberta.ca
Fri Dec 9 14:13:00 EST 1994


In article <3ca71b$lpi at mserv1.dl.ac.uk>, <Enevold.Falsen at alinks.se> wrote:

<Stuff Deleted>

> I suspect an intracellular bacteria or fungi but a virus can 
> not  be excluded completely.

Hmmmm.....

Sorry Jan....this posting sounds a little bit *fishy*....a person in a
ClinViro department asking how to distinguish between a virus and a
bacterium??? 

If this is actually on the up and up.....

Since you claim that the agent can be passaged to uninfected cells it must
be present in the supernatant of infected cultures....take an aliquot from
a culture showing >90% CPE, spin out cells and debris (<1000 g), clarify
sup at 5,000 g for 15 minutes, remove sup for ultracentrifugation at
80-100,000 g for 12-16 hrs. Examine 5K run and ultracent run pellets by
TEM.

Have you tried to create a cDNA from this "novel" 2.5 kb RNA, sequence
it/use it to back-probe DNAs and RNAs from infected and uninfected
cultures?

If I had to make a guess I'd wager it is a ureaplasma/mycoplasma which was
not detected by your Gen-Probe.

Karl

-- 
Karl Fischer
kfischer at gpu.srv.ualberta.ca
tyr-2 at bones.biochem.ualberta.ca



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