[Histonet] West Nile IHC

[Kathleen Jones]

Hello Jan

Could you please post this protocol on histonet, West Nile IHC testing
is something out lab is definitely interested in pursuing. 

Much appreciated, and enjoy the S/C in Seattle!
Kathy




>>> 09/27/10 2:51 AM >>> 
Tresa, 

I've been staining for WNV for many years and we've had a number of
papers 
published with our results. I'm currently at the NSH symnposium, but
will 
forward our SOP to you when I return on Thursday. 

Just a quick note, though. Although we've had many positive results with

birds (esp. raptors), I've only seen one positive cell in all the horses

that I've tried staining. So the fact that you haven't had any positive 
equine tissues does not surprise me. It could be that the horse is an 
end-host of the virus, so there is no replication going on. The virus
might 
be being degraded by cellular processes and thus, not recognized by the 
antibody. 

Jan Shivers 
Section Head 
IHC/Histo/EM 
Veterinary Diagnostic Laboratory 
University of Minnesota 
St. Paul, MN 

On Sep 24 2010, Goins, Tresa wrote: 

After many frustrating attempts, I would greatly appreciate any 
information on the successful application of IHC for the detection of
West 
Nile Virus in fixed tissue. I have tried no retrieval, enzymatic and
heat 
retrieval with no success on horse and bird tissues. 
> 
Would someone please share successful protocols (retrieval) and antibody

sources for equine and avian fixed tissues? Our tissues are formalin
fixed 
and the detection method used is biotin-streptavidin, alkaline
phosphatase 
with a new fuchsin chromogen (Vulcan Fast Red) 
> 
>Thank you in advance, 
> 
>Tresa Goins 
>Veterinary Diagnostic Lab 
>Department of Livestock 
>Bozeman, Montana 
> 
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