Hello,I would like to hear from anyone who is actually doing PCR for Human
Papillomaviruses on DNA from cells obtained by cervical swabs, cytobrush,
cervical lavage etc.  Basically, what works best for you?  For example,
which do you recommend.. brush or swab or?,is the swab, brush etc. left dry
in collection tube?, in a transport media?, at what temperature?  How best
to separate cells?  extraction procedure? and so on. 
Many thanx for any good info.
Dr.Sumeyra kocturk from Turkey

Hello Philipp,

Didi you get any reply on this question. I was too looking for the same.

It would be great if you reply

Thanks

Ekta

Dear Colleague,

I'd like to remind you of the

*VIZIER / SPINE2 Workshop on “Structural Virology*

taking place 14th - 16th July, 2008, Vienna, Austria

see: http://www.projects.mfpl.ac.at/structural-virology/

We can accept up to 100 participants (on a first come first serve
basis) and although we're not there yet the number is starting to
increase.

On behalf of the Organizers,

Paul Tucker

************************************************************************
Dr. Paul A. Tucker

Group Leader

EMBL Hamburg Outstation Tel: +49 40 89902129
c/o DESY, Notkestrasse 85 FAX: +49 40 89902149
D22603 Hamburg Email tucker@embl-hamburg.de
Germany

************************************************************************

Does some one know the molecular weight of Influenza virus proteins (HA,
NP, M, NS etc.) and how do they shown on an SDS-PAGE gel?

Thanks

Sagit

Ziv@biondvax.com

Magda,

Thanks for the correction. I was confusing genome size with virion diameter. Parvovirus is much smaller (approximately 20 nm versus 50 nm).

The low sensitivity of electron microscopy is an issue for clinical diagnosis but should not be for virus grown in culture. I do not have personal experience in quantitation or the accuracy thereof.

I will be interested in data on the accuracy of PFU measurements too. Since it involves serial dilution of a stock the accuracy would depend on how many dilution steps are used before actual plating.

Parmjeet

Parmjeet Randhawa, M.D.
Professor of Pathology,
Division of Transplant Pathology,
University of Pittsburgh, Department of Pathology,
E737 UPMC-Montefiore Hospital,
3459 Fifth Ave, Pittsburgh, PA 15213.
Phone: 412 647 7646
Fax: 412 647 5237

-----Original Message-----
From: virology-bounces@oat.bio.indiana.edu [mailto:virology-bounces@oat.bio.indiana.edu] On Behalf Of Dunowska, Magda
Sent: Sunday, May 11, 2008 5:56 PM
To: Randhawa, Parmjeet; virology@oat.bio.indiana.edu; virology@magpie.bio.indiana.edu
Subject: RE: [Virology] RE: particle to pfu ratio

Magda,

Thanks for the correction. I was confusing genome size with virion diameter. Parvovirus is much smaller (approximately 20 nm versus 50 nm).

The low sensitivity of electron microscopy is an issue for clinical diagnosis but should not be for virus grown in culture. I do not have personal experience in quantitation or the accuracy thereof.

I will be interested in data on the accuracy of PFU measurements too. Since it involves serial dilution of a stock the accuracy would depend on how many dilution steps are used before actual plating.

Parmjeet

Parmjeet Randhawa, M.D.
Professor of Pathology,
Division of Transplant Pathology,
University of Pittsburgh, Department of Pathology,
E737 UPMC-Montefiore Hospital,
3459 Fifth Ave, Pittsburgh, PA 15213.
Phone: 412 647 7646
Fax: 412 647 5237

-----Original Message-----
From: virology-bounces@oat.bio.indiana.edu [mailto:virology-bounces@oat.bio.indiana.edu] On Behalf Of Dunowska, Magda
Sent: Sunday, May 11, 2008 5:56 PM
To: Randhawa, Parmjeet; virology@oat.bio.indiana.edu; virology@magpie.bio.indiana.edu
Subject: RE: [Virology] RE: particle to pfu ratio

Hi everybody,
With respect to Parmjeet's email below: I got the 5.2 from a description in a veterinary virology book, but after your email I did check that info. So, parvovirus genomes are 5 kbp long according to the latest report on virus taxonomy (http://www.ncbi.nlm.nih.gov/ICTVdb/ICTVdB/00.050.1.01.htm) and the available full genome sequences of porcine parvovirus on GenBank are about this length (a link to a recent submission of a whole sequence of 5075 bp: http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=9628826).

Also, I am not sure how accurate the EM would be for quantifying viral particles, as it is a pretty insensitive tool and one needs at least 10*6 particles/mL to be even able to see them under EM, but I would be very interested to know whether or not anybody out there have used it for this purpose?

Magda Dunowska, LW (vet), PhD
Senior Lecturer in Veterinary Infectious Diseases (Virology)
Institute of Veterinary, Animal and Biomedical Sciences
Te Kura Mātauranga Kararehe
Massey University
Palmerston North
New Zealand

Phone : (06) 356-9099 ext 7571
Website : http://ivabs.massey.ac.nz

-----Original Message-----
From: Randhawa, Parmjeet [mailto:randhawapa@upmc.edu]
Sent: Saturday, 10 May 2008 5:43 p.m.
To: Dunowska, Magda; virology@oat.bio.indiana.edu; virology@magpie.bio.indiana.edu
Subject: RE: [Virology] RE: particle to pfu ratio

Could this be a possible treatment for HIV/AIDS:

Synthesize a peptide nucleic acid antisense oligonucleotide for HIV’s
gag gene’s mRNA. The reason for using PNA is that it is substantially
more resistant to enzyme degradation by nucleases and proteases...

I think parvovirus is smaller than 5.2 kb, more like 2-3 kb. Virus
particles could also be quantitated by electron microscopy but that will
not distinguish viable from non-viable virions.

Parmjeet Randhawa, M.D.
Professor of Pathology,
Division of Transplant Pathology,
University of Pittsburgh, Department of Pathology,
E737 UPMC-Montefiore Hospital,
3459 Fifth Ave, Pittsburgh, PA 15213.
Phone: 412 647 7646
Fax: 412 647 5237

-----Original Message-----
From: virology-bounces@oat.bio.indiana.edu
[mailto:virology-bounces@oat.bio.indiana.edu] On Behalf Of Dunowska,
Magda
Sent: Friday, May 09, 2008 7:14 PM
To: virology@oat.bio.indiana.edu; virology@magpie.bio.indiana.edu
Subject: [Virology] RE: particle to pfu ratio

Doria,
If you have a pure virus prep, maybe you could titrate it (to get PFU/mL value), and then extract viral DNA and estimate the number of viral particles based on the amount of viral DNA present, knowing that parvoviruses have a single-stranded DNA genome about 5.2 kbp in size? Just a thought...

Magda

----------------------------------------------------------------------

Message: 1
Date: Wed, 7 May 2008 12:26:10 -0400
From: "Bowers, Doria" unf.edu>
Subject: [Virology] particle to pfu ratios
To: magpie.bio.indiana.edu>
Message-ID:
<783CE756031AB141B53257D89039DD75075EFFA3@DIAMOND.unfcsd.unf.edu>
Content-Type: text/plain; charset="US-ASCII"

We are wondering what the particle:PFU ratio is for porcine parvovirus.

Any ideas how to acquire this statistic?

Doria Bowers

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