[Histonet] RE: Validation - a little detail

Elizabeth Chlipala liz <@t> premierlab.com
Wed Nov 16 12:41:49 CST 2011


When you optimize a stain you are normally just looking at one tissue type or possible a couple, not sure how you handle protocol development.  The amount of work that is required for validation in the clinical setting is dependent upon the type of primary antibody that you are using.  If it is an IVD antibody I believe that verification is only required.  You need to verify that this antibody works as stated in your laboratory.

If you are using an RUO antibody you have go through a validation process.  This in my opinion includes two steps.

1. protocol development and optimization
2. validation - you are checking for sensitivity, specificity and reproducibility on multiple tissue samples

The CAP has a paper that has recommendations for validation (standardization of IHC) for validation they want you to look at 25 different tissue samples, 10 of which are strong positive, 10 which are weak to moderately positive and 5 that are negative.  In the case of prognostic and treatment related IHC they suggest that more tissues are evaluated.  New lots require 3 tissues; strong positive, weak to moderate, and negative.

Ultimately the your laboratory will come up with some system as how you are going to approach this.  This is just a really basic overview and there is a lot more that can be added.  In my opinion I think you need to do what makes sense in your laboratory.  I would have a defined SOP for how you handle certain types of antibodies.

Just my two cents.


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Amber McKenzie
Sent: Wednesday, November 16, 2011 11:21 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Validation

Just wondering what y'alls opinion is on validation:  I don't really understand why optimization isn't enough.  At that point, the pathologist has said what he wanted the stain to look like, so why do 3-10 positive slides on the new instrument to compare to previous slides from another instrument?  Thanks!

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