Re: ER/PR, Her2 Regulatory Questions

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From:Amos Brooks <> (by way of histonet)
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Hi Louri,
    We do ALOT of Herceptest at our lab (between 20 and 40 each day). One
of the
marvels of this test and its stiff regulations about target retrieval is
that it
leaves very little room for variation. You don't (or can't) change the titer,
incubation times or change the retrieval technique. This in it's own complexity
makes valiation and optimization simple for the techs.
    Know in your heart there will always be left over reagent. It is not like
assembling a bookshelf and finding extra screws (been there done that). Your
leftover reagent is still usable next time as long as it is the same lot

    The only people who cant run the test are those who cannot follow the time
constraints to a "T". Reproducibility is the key.
    Another comment worthy of note is that you may not need to run a control
with each case. A control for each run (48 slides) should be sufficient. A
control for each case would burn through all our controls too quick. We run two
slides per test, one with the primary and one with the enclosed negative
reagent. Then for each staining run a positive and negative control and the
enclosed control slide from DAKO.
    Our reps have agreed this is sufficient for the regulations, but extra work
is never frowned upon by inspectors.
Happy Holidays
Amos Brooks

Louri Caldwell wrote:

> Hello everyone,
> I have been asked to initiate immunoperoxidase procedures for ER/PR and Her2
> on breast cancer cases and I want to be sure we follow all applicable
> regulations.  I have read the CAP, FDA, and CLIA guidelines, but I want to
> be sure I've done everything correctly.
> Our current procedure is as follows:  We use Dako's ER & PR primary
> antibodies with their LSAB2 kit for ER & PR, and the Herceptest kit for
> Her2.  The lot of each antibody and kit is tracked as to date
> ordered/received/opened/discarded, along with the initial reactivity of each
> antibody/kit, and the cases tested using each lot.  Along with each case
> tested, a control block is being used that has a range of known reactivity
> to each antibody (from 0 to 3+) and a negative control section of the case
> being tested.  In addition to these 2 controls, the Herceptest kit control
> slide is run with the Her2.  The results of each stain as dictated by the
> pathologist is also tracked.
> Here are the questions I have:
>   Are there any additional controls that need to be run or validation
> procedures I need to go through in order to be compliant?
>   Do I need to go through separate validation procedures if I use less
> reagent than the Herceptest instructions dictate?  If so, to what extent?
>   Are there any restrictions as to who can perform these procedures?
> Any assistance would be greatly appreciated.  Wishing all a very safe and
> happy holiday season.
> Louri Caldwell
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