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Molecular Monitoring in CML: Diagnostic Tests and Treatment Decision Support

Explore laboratory tests like PCR for CML diagnosis & treatment monitoring. Learn about sample collection, analysis methods, & decision-making tools. Discover how PCR & Digital PCR aid in disease monitoring & treatment cessation. Stay informed per European Leukaemia Network Guidelines.

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Molecular Monitoring in CML: Diagnostic Tests and Treatment Decision Support

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  1. Molecular Monitoring in CMLthe process and questions we can answer Letizia Foroni

  2. Aims • Review laboratory tests used for the diagnosis of CML • Review the information provided by PCR • Stopping treatment: how can molecular test help to decide

  3. Process of analysis Sample collection and analysis Sample examination

  4. The first step: look down a microscope! Increased number of one type of cells, the granulocytes o called because they contain a lot of granules!

  5. Other diagnostic tools to confirm the diagnosis? Cytogenetic analysis

  6. Cytogenetics: Philadelphia chromosome Chr 9 Chr 22: Ph chromosome

  7. Philadelphia chromosome

  8. Other diagnostic tools to confirm the diagnosis? Molecular analysis

  9. Sample processing and RNA extraction

  10. PCR analysis

  11. Molecular Monitoring post TKI therapy Are the remaining blood cells normal or leukaemic? For this we require a very sensitive methodology: Real Time PCR or Quantitative PCR

  12. BCR-ABL1: FU analysis Real time quantitative PCR

  13. What information does the PCR test provide?

  14. PCR results

  15. PCR results

  16. PCR and International Scale Values 0.75 Lab 1 Procedure 1 Reference material Lab 2 Procedure 2 Lab 3 Conversion Factor 0.55 Procedure 3

  17. Effect of Conversion factor/s on tests from different laboratories

  18. Monitoring CML summary

  19. European Leukaemia Network Guidelines

  20. When is it important to test? • At diagnosis • Every 3 months for at least the first 2 years (general practice) but to be discussed with your clinical team. • Every 6 months after 2 years.

  21. Monitoring CML summary CAN we stop?

  22. The best method to use to decide when is best to stopWhich and when?

  23. Digital PCR

  24. Conclusions • PCR provides the most sensitive method to monitor disease • Samples at 3 months and every three months predict future outcome • Digital PCR is helping to identify patients who can consider stopping therapy

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