RE: Scrapie infected tissues

From:"Monson, Frederick C."

http://www.who.int/emc/diseases/bse/bsecjd.html


From CDC:  BMBL Section VII Agent Summary Statements
Section VII-D: Prions
http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4s7d.htm

Click on "Table 5......" in left frame.  Extracted below BUT you should read
all at site!!!!!!

Table 5. Tissue preparation [in/from autopsy]

1. Histology technicians wear gloves, apron, laboratory coat, and face
protection. 

2. Adequate fixation of small tissue samples (e.g. biopsies) from a patient
with suspected prion disease is followed by post-fixation in 96% absolute
formic acid for 30 minutes, followed by 48 hours in fresh 10% formalin. 

3. Liquid waste is collected in a 4L waste bottle containing 600 ml 6N
sodium hydroxide. 

4. Gloves, embedding molds, and all handling materials are disposed of as
biohazardous waste. 

5. Tissue cassettes are processed manually to prevent contamination of
tissue processors. 

6. Tissues are embedded in a disposable embedding mold. If used, forceps are
decontaminated. 

7. In preparing sections, gloves are worn, section waste is collected and
disposed of in a biohazard waste receptacle. The knife stage is wiped with
1-2N NaOH, and the knife used is discarded immediately in a "biohazard
sharps" receptacle. Slides are labeled with "CJD Precautions." The sectioned
bloc is sealed with paraffin. 

8. Routine staining: 

a. Slides are processed by hand. 

b. Reagents are prepared in 100 ml disposable specimen cups. 

c. After placing the coverslip on, slides are decontaminated by soaking them
for 1 h in 2N NaOH. 

d. Slides are labeled as "Infectious-CJD." 

9. Other suggestions: 

a. Disposable specimen cups or slide mailers may be used for reagents. 

b. Slides for immunocytochemistry may be processed in disposable petri
dishes. 

c. Equipment is decontaminated as described above. 

THE ABOVE MAY INFORM BUT IT CANNOT COMFORT.  THESE AGENTS ARE EXTREMELY
DANGEROUS AND PROBABLY SHOULD NOT BE APPROACHED VOLUNTARILY IN A SITUATION
WHERE BIOSAFETY LEVEL IS BELOW III (3).  CLEANING UP AFTER HISTOLOGIC
PREPARATIONS WOULD DISSUADE ME.

Regards,

Fred Monson

P.S.  I have experience with sterility, but not with agents like these,
thank goodness!

Frederick C. Monson, PhD   
Center for Advanced Scientific Imaging
Schmucker II Science Center
c/o Geology/Astronomy
West Chester University
South Church Street and Rosedale Ave
West Chester, Pennsylvania, USA, 19383
Phone:  610-738-0437
FAX:  610-738-0437
fmonson@wcupa.edu
CASI URL:  http://darwin.wcupa.edu/casi/
WCUPA URL:  http://www.wcupa.edu/
Visitors URL:  http://www.wcupa.edu/_visitors/


> ----------
> From: 	Histo-Scientific Research Laboratories
> Sent: 	Wednesday, September 4, 2002 2:59 PM
> To: 	Histonet
> Subject: 	Scrapie infected tissues
> 
> Dear Netters,
>  
> We have been asked to prepare slides from scrapie infected tissues.  I
> have been able to find a couple of abstracts regarding this disease but we
> are still skeptical about processing these tissues with our routine
> specimens.  Has anyone worked with this disease?  If so, what precautions
> were taken (other than the normal precautions)?  I have been told by a
> researcher from the USDA that formalin does not stabilize the disease.
> However, I have also been told that formic acid or sodium hydroxide will
> fix/stabilize the tissue.  I would appreciate any information from someone
> who has worked with this disease.
>  
> thanks,
>  
> Tom Galati
> Laboratory Director
> HSRL-A GLP Compliant Laboratory
> 137 S. Main Street
> Woodstock, VA  22664
> www.hsrl.org
> tomgalati@hsrl.org
> (540)459-8211
> fax: (540)459-8217
> 



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