Molecular Monitoring of Chronic Myeloid Leukemia

Methods Mol Biol. 2020:2065:153-173. doi: 10.1007/978-1-4939-9833-3_12.

Abstract

Molecular diagnosis and measurement of minimal residual disease (MRD) in patients with chronic myeloid leukemia (CML) is essential for clinical management. In the era of tyrosine kinase inhibitor therapy molecular tests including BCR-ABL1 transcript monitoring and kinase domain mutation analysis are the main tools used to inform choice of treatment, appropriate dosage and even whether therapy can be safely withdrawn. Quantitation of BCR-ABL1 oncogene transcript by real-time quantitative PCR (qPCR) is currently the gold-standard method for monitoring as it provides superior sensitivity over karyotyping and fluorescent in situ hybridization (FISH). Here we describe step-by-step methods of RNA conversion to cDNA along with the qPCR protocol which is used in one of the main reference laboratories for this test.

Keywords: BCR-ABL1; CML; Multiplex PCR; TaqMan; cDNA synthesis; qPCR.

MeSH terms

  • Bone Marrow / pathology
  • Fusion Proteins, bcr-abl / antagonists & inhibitors
  • Fusion Proteins, bcr-abl / genetics
  • Gene Expression Profiling / methods*
  • Humans
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / blood
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / diagnosis*
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / drug therapy
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics
  • Multiplex Polymerase Chain Reaction / methods*
  • Neoplasm, Residual
  • Protein Kinase Inhibitors / pharmacology
  • Protein Kinase Inhibitors / therapeutic use
  • RNA, Messenger / genetics
  • RNA, Messenger / isolation & purification
  • Real-Time Polymerase Chain Reaction / methods*

Substances

  • BCR-ABL1 fusion protein, human
  • Protein Kinase Inhibitors
  • RNA, Messenger
  • Fusion Proteins, bcr-abl