[Histonet] IHC validation

BSullivan <@t> shorememorial.org BSullivan <@t> shorememorial.org
Thu Feb 10 06:21:52 CST 2011

Added to this.....................make sure you keep your work up sheets.

Beatrice Sullivan, HT(A.S.C.P.) HTL , AAS, CLSP(N.C.A.)
AP Supervisor
Shore Memorial Hospital

Speak only well of people and you need never whisper

             Rene J Buesa                                                  
             <rjbuesa <@t> yahoo.co                                             
             m>                                                         To 
             Sent by:                  Histonet                            
             histonet-bounces@         <histonet <@t> lists.utsouthwestern.edu> 
             lists.utsouthwest         , Joe Nocito <jnocito <@t> satx.rr.com>  
             ern.edu                                                    cc 
             02/09/2011 03:26          Re: [Histonet] IHC validation       

This is what I would do;
1- run 1 control slide per antibody you have in your "arsenal"
2- compare the result with a control slide already in your files.
3- show both slides to the chief pathologist (after all is his/her opinion
the one is going to be asked by CAP)
4- those antibodies whose positive controls reacted substantially different
to those in your files are the ones you have to work with with respect to
concentration or detection method.
5- never overdue it, and avoid excessive costs that usually are never
Rely always in your pathologist's opinion
René J.

--- On Tue, 2/8/11, Joe Nocito <jnocito <@t> satx.rr.com> wrote:

From: Joe Nocito <jnocito <@t> satx.rr.com>
Subject: [Histonet] IHC validation
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Date: Tuesday, February 8, 2011, 5:43 PM

Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for specificity.
Hopefully we can obtain cases that are really positive, some weakly
positive and some flat out negative. Once that is completed, we run 10
different tissue types to check for any unexpected cross-reactivity.  
The ultra holds 30 slides and we are receiving two machines. If we run 10
slides/antibody, that's going to take a while, not to mention the number of
detection kits that will be used. Do you think 5 slides/antibody is
sufficient? I emailed CAP last week for their take and they never returned
my email (I told my medical director to hold their check for the year and
see how fast they respond to that). Ah oh, don't go down that road Joe,
it's unhealthy. What are your thoughts? Thanks

Joe (JTT)

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