[Histonet] IHC validation

Rene J Buesa rjbuesa <@t> yahoo.com
Wed Feb 9 14:26:55 CST 2011

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 appreciated.
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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