Re: CAP Ploidy and Cell cycle Survey and comments

Colleagues:
this seems to be a serious issue that is not resolved. I would 
like to see this addressed in some kind of official manner 
that might result in a resolution. Frankly, this is one of the 
things that ISAC should do and I would like to propose that 
this is taken up by ISAC immediately.

If there are a number of people that would like to particpate 
in some kind written evaluation of these surveys, I will 
happily facilitate it. I am very concerned that there appears 
to be some discontent in standards and QC evaluations. So, 
if I can receive suggestions of what you would like to see, i 
think that we can more officially address this from the 
position of ISAC. At least we can try???

Kind regards
paul robinson
President-elect, ISAC

On 17 May 2005 at 10:29, Tom Sawyer wrote:

> Hi Tim - My contribution........ don't know how lively it is.
> 
> 1. I feel stupid re #1. I initially reviewed the participant results and saw we
> did well with a DI = 1.4. Didn't pay attention to what THEY said we should get.
> Obviously they either sent the incorrect sample or were given the incorrect
> information as to what the DNA content of Daudi cells actually is. I've never
> worked with this cell line so I don't know what the "correct" answer is. It
> really is amazing that they did not comment at all on this discrepancy. I had a
> similar experience with CDC a while back on an MPEP survey. They send a
> preliminary report (what they got) and then a report of participant results. On
> one sample we were way off of their result but when the participant report came
> out we were perfect. I called and got a similar response to your response from
> CAP. They said, "gee, I guess we should have commented on our mistake in the
> final report." We all make mistakes and should be forthcoming when we do,
> including the surveying agents.
> 
> 2. I cannot comment on your perception that all clinical labs do paraffin
> embedded samples except to say we only do fresh blood, bone marrow and tissue
> samples in our lab. My problem with the CAP samples is that they are
> acid-alcohol fixed which we do not do. All our real samples are processed using
> the Vindelov method and we get full peak CV's in the range of 1.2-1.4% on our
> PBMC controls and usually less then 2% on our patient samples. We therefore can
> barley resolve DNA content aneuploid cells with DI's in the range of 1.03-1.04.
> The best we've been able to do on the CAP calibrator is in the range of
> 3-4%.This seems to be about the mean for the group on this sample. Given this,
> it would be ALMOST IMPOSSIBLE for the AVERAGE participant to resolve two peaks
> when the DI is in the ranges specified for this shipment 1.07-1.09. WHY WOULD
> THE PERSON RESPONSIBLE FOR SELECTING SAMPLES SEND OUT MATERIAL THAT WOULD BE
> STATISTICALLY PREDICTED TO FAIL?
> 
> 3. I agree with you that our surveys (in the USA) are not what we would like
> them to be. The only good one is the CDC MPEP which sends real patient samples.
> The CAP survey for Lymphocyte Immunophenotyping has been significantly
> diminished since they began sending StausFlows. These fixed cells do not behave
> the same as patient samples. We have to use completely different instrument
> settings because of the high autofluorescence. Immunophenotyping cell lines for
> leukemia and lymphoma has never made much sense to me and has little bearing on
> what we do on a daily basis in our laboratories; but we all saw what happened
> with spiked blood samples. I really don't know the answer to this one.
> 
> 
> Now that I've slammed CAP I must say that I empathize with the magnitude of the
> problem. Everyone in clinical cytometry knows that we all get the best results
> on fresh specimens. It must be a daunting task to be able to prepare, package,
> and ship samples all over the country and have them arrive in a state that
> people can still work with. I suppose it would be far worse to have real sample
> that no one could analyze.
> 
> Thanks for bringing this subject up,
> 
> Tom
> 
> Tom Sawyer
> Flow Cytometry Specialist
> Flow Cytometry Laboratory 32HE
> Medical College of Ohio
> 3055 Arlington Ave.
> Toledo, OH  43614-5806
> Phone:	419.383.4212
> Fax:	419.383.3076
> email:	tsawyer@mco.edu
> 
> 
> >>> "Tim Kute" <tkute@wfubmc.edu> 05/11/05 10:57 AM >>>
> Dear All,
> 
>     I recently got back our results from the CAP survey to measure
> ploidy and S phase activity.	This survey is used for quality control
> for hospital facilities that do
> 
> flow cytometry analysis.  It costs $681/yr and there are 4 tests per
> year.  I have talked to CAP and my comments are listed below:
> 
> 
> 1.  Very little comments are given back.  Interesting in the last
> survey, the FL-06 was reported to be aneuploid ( DI= 1.07), 30%
> aneuploid cells and cell cycle was 16%. The results from the surveyed
> group including us were DI of 1.42, 26% aneuploid cells, and S% of 27.3.
> Why was this different never discussed?
> 
> 2.  Why is CAP still using cell lines when I would believe that all
> clincal samples are from paraffen embedded material?	The CAP said that
> they did a survey and found that most labs were happy to have it the way
> it is now and therefore CAP has decided not to change it.  In my
> opinion, the CAP should be making the decisions and not the hospitals
> who are wanting easy tests that make them look good.
> 
> 3.  The US is behind the times in quality control.  If you read the
> literature, Europe has a well defined quality control flow cytometry
> program and uses frozen samples for their studies.  They have never got
> into studies with paraffin embedded material but at least this better
> tests the lab procedures than what is currently being done by CAP.  
> 
> 
> I would be interested in a lively discussion of these issues and I will
> try to give a summary after they are prsented.
> 
> 
> Dr. Tim Kute
> 
> tkute@wfubmc.edu 
> 
> 
> 


J.Paul Robinson, PhD		 PH:(765)4940757
Professor of Immunopharmacology
Professor of Biomedical Engineering  
Purdue University	   FAX:(765)4940517
EMAIL:jpr@flowcyt.cyto.purdue.edu
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