RE: A big thank you on biosafety

Dear Joern,

 

Apparently it only went out to you.

Re how it was validated. I'm not really sure. I wasn't involved in the
whole risk assessment exercise we went through as I was only a Research
Assistant at that time, but we were told to use bleach to clean the
benches or (not sure about the exact conc here) 10 min 4M Hydrochloric
acid for re-useable scalpels and plasticware. And that was in relation
to potential BSE containing cow brain material (which is potentially
infectious to human and handled in a 2b Hazard environment). Also
coveralls and such were autoclaved twice for 30min at 121 degree Celsius
before put in the normal waste.

 

The hazard for non-BSE material such as moose and elk is much lower, but
on the other hand I do subscribe to "better safe than sorry". Not sure
if it has to be undiluted bleach - I guess that's not too good for the
lines in a FACS machine. 

 

Petra 

 

-----Original Message-----
From: Joern Schmitz [mailto:jschmitz@bidmc.harvard.edu] 
Sent: 17 September 2006 03:20
To: cyto-inbox
Cc: 'Cytometry Mailing List'
Subject: RE: A big thank you on biosafety

 

Dear Petra, I don't know whether your response came only to me or really
went out to everybody in the flow Cytometry group. 

 

Just to comment on your e-mail: I think all of this is a very healthy
and urgently needed discussion to ensure all cell sorter operators are
reasonably well protected when they perform cell sorts of potentially
biohazardous specimens. 

 

When I read your comment at the bottom of your e-mail: "put some
undiluted bleach on it" a number of questions come up. Just to list a
few: 

1.    How has that been validated?

2.    Do we truly need to use undiluted bleach? For how much time?

 

As I am not directly working in this field (however oversee a flow
cytometry facility that sorts AIDS virus-infected nonfixed human and
non-human primate specimens), I leave the rest of the discussion to the
experts in this field. 

 

Regards, Joern

 

Joern E. Schmitz, MD

Assistant Professor of Medicine

Harvard Medical School

 

Division of Viral Pathogenesis

Department of Medicine	     

Beth Israel Deaconess Medical Center

Research East 213D

41 Avenue Louis Pasteur

Boston, MA  02115

 

Phone: 617-667-5206

Fax:   617-667-8210

 

http://bidmc.harvard.edu/display.asp?leaf_id=4102

 

http://www.hms.harvard.edu/aids/programs/cfar/cores/immflowlma.htm

 

This document may contain information that is privileged, CONFIDENTIAL
and exempt from disclosure under applicable law. If you are not the
intended recipient, please notify me immediately, as the use of this
information is strictly prohibited.

 

-----Original Message-----
From: Disterer, Petra (Medsch Hampstead/Medicine)
[mailto:p.disterer@medsch.ucl.ac.uk] 
Sent: Saturday, September 16, 2006 6:20 AM
To: cyto-inbox
Subject: RE: A big thank you on biosafety

 

Dear Flowers,

 

Having worked with Prions for three years, I think I should put my two
cents in. There is no evidence at all that TSE from elk or moose is
transmissable to human. On the other hand there is considerable evidence
that cross-species TSE infection is extremely difficult, for example
infecting mice requires high-dose administration of pure beta sheet
prion protein directly to the brain and passaging into other mice.
Scrapie also is impossible to transmit to human and it's been around for
hundreds of years, so we really would know by now.

 

So basically I would estimate the risk to be very low. For safety
precaution I would see if the cells could be fixed before sorting and
avoid aerosol formation as you would with any potentially infectious
sample, but I'm just suggesting that for your peace of mind. I as the
researcher working with those cells would not take any precautions
beyond what I would usually with all tissue culture.

 

I'm assuming here that with "material" you meant cells and not purified
prion protein. I don't think you could FACS that, as the sizes wouldn't
be big enough.

 

Again for line purification - you'd be coping with cells that
potentially contain prion protein not free concentrated prion protein,
so I think that cleaning according to what you normally do for
potentially infectious cells would be adequate.

 

Just to mention, prion protein (the normal alpha helical form and
potentially some misfolded forms) are present in quite a few cells which
you've probably already sorted, such as central nervous sytem neuronal
cells or Purkinje cells.

 

I think we as scientists should be careful about subscribing to the
media induced panic about prion protein. (Ie prion protein is not
practically indestructable - put some undiluted bleach on it and that'll
do fine).

 

Hope that helps

 

Petra Disterer

The UCL Inst.of Hepatology

 

      -----Original Message----- 

      From: Joern Schmitz [mailto:jschmitz@bidmc.harvard.edu] 

      Sent: Fri 08/09/2006 22:05 

      To: Cytometry Mailing List 

      Cc: 

      Subject: RE: A big thank you on biosafety

      

      

 

      I totally agree: The Institutional Biosafety Committee has to look
at this.

      

      BTW: Who has decided that the biosafety level should "only" be
BSL-2 when

      you are handling "bugs" that are basically indestructible?

      

      My gut feeling tells me that right now nobody really has any clue
about any

      potential longterm effects of aerosols from these specimens that
you are

      about to generate ...

      

      Joern E. Schmitz, MD

      

      -----Original Message-----

      From: Charles A Kuszynski [mailto:ckuszyns@UNMC.EDU]

      Sent: Thursday, September 07, 2006 11:22 AM

      To: cyto-inbox

      Subject: Re: A big thank you on biosafety

      

      I would be more concerned about the potential for aerosol
production and

      biosafety issues than whether you can clean the sample tube.  Your

      institutional Biosafety Committee needs to look at this project
and suggest

      the requirements for operator safety and containment.

      

      Charles A. Kuszynski, Ph.D.

      

      

      Hello,

      

      An investigator here has inquired about using the MoFlo for
sorting

      prion infected material.	This will be from deer and elk with
chronic

      wasting disease, which is treated as BSL2, and may eventually also
be

      scrapie, which I think is also BSL2.

      

      I was wondering if anyone out there has done these kinds of sorts,
and

      if so, what sort of containment and line decontamination they have
used?

      

      Thanks very much.

      

      Anne

      

      

 


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