Quantification of virus particles - followup

From: Reed, Doug S Dr USAMRIID (Doug.Reed@DET.AMEDD.ARMY.MIL)
Date: Tue Apr 04 2000 - 11:59:45 EST


Thanks to everyone who has responded so far (and to those who might still
respond). I think the best solutions I've seen so far were Howard Shapiro's
solution of using anti-vaccinia antibodies (PE conjugated) and (even better,
although technically more difficult) putting various pfu concentrations of
virus under an electron microscope and counting.

Some of you commented that you weren't sure that the pfu/particle ratio
wasn't that important. At this point it is unknown what effect it has - but
we have routinely seen viable bacteria counts by flow that are 1-2 logs
higher than the colony counts after aerosol sprays. At this point it is
interesting information, but the relevance is unclear.

Howard - you are right, we could count vaccinia directly by flow given the
large size of poxviruses. I am attempting to develop the beads as a more
generally adaptable technique to the wide variety of viruses we study here -
not just vaccinia but also the more exotic filoviruses which are
considerably smaller. Currently most researchers use vero plaque assays to
determine the pfu of the virus, which is certainly a proven technique but
requires considerable time and effort whereas the beads could be done the
same afternoon as the spray.

Many thanks again to all those who replied!

Douglas S. Reed, Ph.D.
Microbiologist
Department of Aerobiology and Product Evaluation
Division of Toxinology and Aerobiology
U.S. Army Medical Research Institute of Infectious Disease
1425 Porter St. Ft. Detrick
Frederick, MD 21702-5011
301-619-6728
301-619-2541 fax
Doug.Reed@det.amedd.army.mil



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