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Chronic Myeloid Leukemia: a Case Study- only apply if you are familiar with the topic as well as PCR and DNA sequencing!

| October 30, 2015

I will soon upload a file named “Methods and Results New”; please write a discussion/conclusion for the paper in the section following the heading ‘Discussion’.
Please, I want you to first briefly tell me what you can tell from the results. So you’ll need to read the methods and results sections THOROUGHLY. (Note: a good response=1 log reduction by 1month, 2 log reduction by 3 months, 3 log reduction by 6 months. A complete molecular response CMR= 0 BCR-ABL signal). Discussion should include reference to the database of known ABL mutations (I’ll upload this shortly) and we need to make recommendations for future treatment regimens appropriate for each patient. Conclusion should refer to the potential of molecular methods to aid in the diagnosis, monitoring, and mutation detection for personalized medical treatment.

This is what I can tell for now:
1. end-point PCR: The most common breakpoints for BCR-ABL are B2A2 and B3A2. This has been well-documented in many research papers and review articles (please find a reference article for this; I only know this because our professor told us so), and our results support this. Of the 20 patients we analysed, all except 2 patients had either B2A2, or B3A2, or both (there were 2 patients who had both). I’m not sure what we should say about the 2 patients who did not have any of these, or the 2 patients who had both. Well, I initially thought that (1) the 2 patient could have had mutations in the primer binding sites such that the primers could no longer bind to their normal binding sites….but this would have resulted in other problems unless the mutation was a silent one. And the chance of having a mutation at the primer binding sites as well as the chromosomal aberration is low, I think. (2) or it could have been that the patient had a different rare mutation that does not involve BCR exon 13,14 or ABL exon 2,3. I think this is more likely. If you look up google for “CML breakpoints”, you should be able to see what I mean by this. (3) or it could have been just due to experimental errors: too much cDNA was lost during the procedure, or too little was loaded onto the gel that it ended up undetectable, or it was mistakenly not added. Can you think of any other experimental errors? Please tell me if you can come up with a different explanation, or which one you think is most likely.
For
2.Qualitative PCR & Gene sequencing: Patient 1 showed only a sub-optimal response to Imatinib although no mutation was found with DNA sequencing. Furthermore, the BCR-ABL/BCR ratio slightly increased in the latest month.The patient needs to be monitored regularly after this, because it is very likely that the patient will soon develop a mutation that confers resistance to Imatinib. In that case, depending on the nature of the drug, the patient will need to receive either an increased dose of Imatinib or another drug such as Dasatinib. I’ll write more in a personal msg.

Category: Biology

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