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