[Histonet] West Nile IHC

shive003 <@t> umn.edu shive003 <@t> umn.edu
Mon Sep 27 01:48:38 CDT 2010


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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