Question about CML patients and doubts about chromosome 20q

[celbio]

Hi everyone,

I have two questions concerning our leukemia patients.

 

1. Since januari this year we receice bonemarrow  from a new hematologist/oncologist and we can not find any chromosome abnormality !

    Even for the CML patients we do not find the philadelphia chromosome; t(9;22). This is very strange, because the years before in almost every CML request we found

    the philadelphia with conventional GTG-banding; even in complex rearrangements (t(1;9;22) and t(2;9;22)). 

    The big difference between de the patients of this year and the years before is the number of leuco's.

    The bonemarrows sent in by the new hematologist/oncologist have leuco's between 10 - 50 and the bonemarrows sent in by the other hematologist/oncologists had much

    higher  number of leuco's > 100.

    I was wondering if this could be the reason why we have not found the philadelphia chromosome still. Even with the FISH  we do not find the t(9;22).

    We still use the same culturemedia and protocols for setting up and harvesting.

    Does anyone have any suggestion for our findings ?

 

2. The next problem is a patient with aplastic anaemie, DD : MDS.

    Echo abdomen shows no splenomagalie.  

    Hb:8.4, mcV: 84, Leuco's: 3.4, thrombo's: 49 and retic: 15.

    We analysed 20 metaphases and in 4 cells we have some doubts about chromosome 20q. See the foto's attached.

    We do not have the FISH probe for del(20q), so we unfortunately can not get rid of this doubt.

    Could anyone please have a look and have a comment on the chrom 20 ?

    And what do you advice us to give as a result?

 

Thank you so much for your help,

Celbio

[dranuraggupta30]

Hello Celbio,

In regards to your first question

1. Please check if the patient is taking some medication like imatinib or desatinib or something else and for how long as if the patient really is a case of CML, he can go into cytogenetic remission after starting medications

2. If the patient is a fresh case of CML then there could be a cryptic t(9;22) for which u may do FISH using  BCR ABL probe which will confirm or refute the diagnosis and if positive its a case of cryptic t(9;22). In 1-5% of the CMLs the translocation may be cryptic.  

3 If ur hospital has a molecular lab then see if u get a BCR ABL transcript by PCR in the bone marrow sample. Its presence or absence will give you the answer.

With high leuco counts chances of culture failure is quite high but not absent Ph.

regarding your second question,

The patient does have cytopenias bicytopenias definitely but pan questionable as ANC is not mentioned. Besides this the MCV is not raised and the retics are high so chances of it being an MDS is quite low.

However looking at your karyotypes, the 20s looks very fine no del (20q) for sure and usually in del (20q) which usually is associated with a very good prog in MDS, the derivative chromosome 20 becomes significantly short which is not the case here. So i thinkk you can report it as a normal karyotype.

Please let me know history of the patient and if u do FISH on the first case or any PCR findings

regards.

[Anurag Gupta]

Hello Celbio,

I regards to your first question

1. Please check if the patient is taking some medication like imatinib or desatinib or something else and for how long as if the patient really is a case of CML, he can go into cytogenetic remission after starting medications

2. If the patient is a fresh case of CML then there could be a cryptic t(9;22) for which u may do FISH using  BCR ABL probe which will confirm or refute the diagnosis and if positive its a case of cryptic t(9;22). In 1-5% of the CMLs the translocation may be cryptic.  

3 If ur hospital has a molecular lab then see if u get a BCR ABL transcript by PCR in the bone marrow sample. Its presence or absence will give you the answer.

With high leuco counts chances of culture failure is quite high but not absent Ph.

regarding your second question,

The patient does have cytopenias bicytopenias definitely but pan questionable as ANC is not mentioned. Besides this the MCV is not raised and the retics are high so chances of it being an MDS is quite low.

However looking at your karyotypes, the 20s looks very fine no del (20q) for sure and usually in del (20q) which usually is associated with a very good prog in MDS, the derivative chromosome 20 becomes significantly short which is not the case here. So i thinkk you can report it as a normal karyotype.

PLease let me know history of the patient and if u do FISH on the first case or any PCR findings

regards.

--
You received this message because you are subscribed to the Google Groups "Cytogenetics methods and trouble-shooting Forum" group.
To unsubscribe from this group and stop receiving emails from it, send an email to cytogenetics-methods-and-t...@googlegroups.com.
To post to this group, send an email to cytogenetics-methods...@googlegroups.com.
Visit this group at http://groups.google.com/group/cytogenetics-methods-and-trouble-shooting?hl=en-GB.
For more options, visit https://groups.google.com/groups/opt_out.
 
 

--

Anuraag gupta 

Dr. Anurag Gupta (MBBS, MD Path, PGDHHM)

Fellow Cytogenetics

Dept. Of Cytogenetics

Tata Medical Center Trust

New Town, Kolkata

9831017344

[Pat Cammarata]

Hi,

 

Question 1. Did you see what the pathology  and flow cytometry  results were for these patients? Perhaps they were normal and not CML as the new doc suspected. Also, were the specimens clotted? We just did a review of over 200 specimens regarding specimen quality and found a dramatic decrease in the abnormal rate if the specimen was clotted. Even if the specimens were diluted with peripheral blood, you should still be able to find some abnormals if the patients had some immature cells in peripheral blood.

Question 2. I think the 20's are normal.

 

Pat

---

--

[celbio]

 

 

Thank you so much for your comments.

 

First of all, all patients are fresh CML cases and we also did the FISH with the bcr/abl probe dc df (from Cytocell) on two of them, after the doc said she was sure that they 

had CML. For both patients we did not find the t(9;22) with the FISH. 

Unfortunately, we do not have pcr for bone marrow in our institute here in Surinam.

 

The second thing is that we indeed received a few bone marrows which were clotted. We immediatly tried to "dissolve" the clott with a needle and transfer some of the

dilluted bone marrow into the tubes with culture media. We have always got a reasonable amount of metaphases doing it this way (only a few times with very low yield; as suspected).

It is good to know that the yield of abnormal cells can be much lower in samples which are clotted ! 

So, I am wondering how we can avoid the bone marrow to get clotted. We receive the bone marrows in a lithium heparine tubes.

 

Concerning the patients with high leuco counts: when this is the case we seed with less ml bone marrow and we always get/got good yield.

And it is in these patients that we indeed find the t(9;22).

In the recently received patients, the number of leuco's is much lower, so we seed with more bone marrow and we get good yield, but we do not find the t(9;22).

Even with the FISH we do not find the t(9;22).  So we wonder if we why we do not find abnormal clones these days.

Do I understand it correct: when we have good yield (despite clotts or high leuco counts) and if the t(9;22) clone is present, we would always find it?

Ofcourse sometimes FISH is needed.

Again, if we have enough metaphases we should be able to find an abnormal clone if present?

 

Does anyone know good literature about the leukemias. I want to now for example how often chromosome abnormalities are found in the different types of leukemias?

I have the book of Rooney, Human Cytogenetics; malignancy and acquired abnormalities, third edition.

 

 

Thanks again,

Celbio

 

 

 

 

Op zaterdag 18 mei 2013 12:32:47 UTC-3 schreef celbio het volgende: