Molecular Monitoring in CML the process and questions we can answer Letizia Foroni
Jan 29, 2016
Molecular Monitoring in CML
the process and questions we can
answer
Letizia Foroni
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
Process of analysis
Sample collectionand analysis
Sample examination
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!
Other diagnostic tools to confirm the diagnosis?
Cytogenetic analysis
Chr 9
Chr 22: Ph chromosome
Cytogenetics: Philadelphia chromosome
Philadelphia chromosome
Other diagnostic tools to confirm the diagnosis?
Molecular analysis
Sample processing and RNA extraction
PCR analysis
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
BCR-ABL1: FU analysisReal time quantitative PCR
What information does the PCR test
provide?
PCR results
PCR results
ConversionFactor
PCR and International Scale Values
Lab 1
Lab 2
Lab 3
Procedure 1
Procedure 2
Procedure 3
Reference material
0.75
0.55
Effect of Conversion factor/s on tests from different laboratories
Monitoring CML summary
European Leukaemia Network Guidelines
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.
Monitoring CML summary
CAN we stop?
The best method to use
to decide when is best to stop
Which and when?
Digital PCR
ConclusionsConclusions•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