[Histonet] MMR IHC question

Sue Hunter SHUNTER <@t> beaumont.edu
Fri Jun 7 07:16:42 CDT 2013

Hi Curt
I agree with Mark - you can set up lab developed tests or - depending on your volumn, experience, requirements for FDA approval, etc - you can buy kits.
We often reflex to MSI testing - run on capillary electrophoresis instruments- we have ABI-310s.  Our primer sets are lab developed but you can also buy them.   We also do KRAS, BRAF, EGFR(mostly on lungs), and MLH1 methylation by real time on an ABI 7500.  Easy to set up and run and fast run time.  Kits are from EntroGen, MLH1 is lab developed.  We do manual DNA extractions using the Qiagen extraction kits.  
Hope this helps.

Sue Hunter, Supervisor
Advanced Diagnostics
Beaumont Health System
Royal Oak MI
shunter <@t> beaumont.edu

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Mark Tarango
Sent: Thursday, June 06, 2013 1:55 PM
To: Curt
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] MMR IHC question

Hi Curt,

You can setup these assays from scratch if you know what you're doing, but if you want FDA approved or to buy simple kits then you need to go with a company that sell that.  Roche just recently had their EGFR assay FDA approved.

We are using Qiagen products for EGFR, KRAS, BRAF.  We use the QIAcube for DNA extraction, the QIAgility for liquid handling in preparation of master mixes, and the rotorgene for real-time PCR.  There are different tests to reflex to at the  point of MMR negative IHC staining.  Some labs do BRAF and/or KRAS as a surrogate marker for sporadic CRC.  Other labs might do methylation testing directly.  We're currently validating a sequencing methylation assay for MLH-1 using Qiagen's pyrosequencing platform (PyroMark).  You should definitely have a pathologist involved in this.


On Thu, Jun 6, 2013 at 9:23 AM, Curt <c.tague <@t> pathologyarts.com> wrote:

> Hello histo-world,
> This might not exactly be a histo question but I know you are all so 
> knowledgeable, it couldn't hurt to ask.
> I'm looking to add some molecular testing to our menu, specifically 
> reflex testing when there is an absence of a MMR protein, let's use 
> Lynch syndrome as an example. First there is a pane of IHC (MLH1, 
> MSH2, MSH6, PMS2), depending on the results the next step is reflex for molecular tests.
> My simple question is this, what type of equipment are these molecular 
> test run on? Are there universal platforms that run molecular tests 
> for a variety of tumors, the only variation would be the "kit" for 
> that specific gene or set of genes?
> Does that make sense, what platform or machine is needed to run 
> molecular tests?
> Thanks,
> Curt
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