A novel RT-qPCR assay for quantification of the MLL-MLLT3 fusion transcript in acute myeloid leukaemia

Eur J Haematol. 2013 Nov;91(5):394-8. doi: 10.1111/ejh.12156. Epub 2013 Oct 1.

Abstract

Objectives: Patients with acute myeloid leukaemia (AML) of the monocytic lineage often lack molecular markers for minimal residual disease (MRD) monitoring. The MLL-MLLT3 fusion transcript found in patients with AML harbouring t(9;11) is amenable to RT-qPCR quantification but because of the heterogeneity of translocation break points, the MLL-MLLT3 fusion gene is a challenging target. We hypothesised that MRD monitoring using MLL-MLLT3 as a RT-qPCR marker is feasible in the majority of patients with t(9;11)-positive AML.

Methods: Using a locked nucleic acid probe, we developed a sensitive RT-qPCR assay for quantification of the most common break point region of the MLL-MLLT3 fusion gene. Five paediatric patients with t(9;11)-positive AML were monitored using the MLL-MLLT3 assay.

Results: A total of 43 bone marrow (BM) and 52 Peripheral blood (PB) samples were collected from diagnosis until follow-up. Two patients relapsed, and both were MRD positive in BM after first induction course. A total of three relapses occurred, and they were detected by RT-qPCR 3 wks before haematological relapse was diagnosed.

Conclusion: This MLL-MLLT3 RT-qPCR assay could be useful in MRD monitoring of a group of patients with AML who often lack reliable MRD markers.

Keywords: MLL-MLLT3; acute myeloid leukaemia; minimal residual disease; RT-qPCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / therapeutic use
  • Biomarkers, Tumor / genetics*
  • Bone Marrow / chemistry
  • Child
  • Child, Preschool
  • Chromosomes, Human, Pair 11
  • Chromosomes, Human, Pair 9
  • DNA Probes / chemistry
  • DNA Probes / genetics
  • Humans
  • Infant
  • Infant, Newborn
  • Leukemia, Myeloid, Acute / diagnosis*
  • Leukemia, Myeloid, Acute / drug therapy
  • Leukemia, Myeloid, Acute / genetics
  • Leukocytes, Mononuclear / chemistry
  • Neoplasm, Residual / diagnosis*
  • Neoplasm, Residual / drug therapy
  • Neoplasm, Residual / genetics
  • Oligonucleotides / chemistry
  • Oligonucleotides / genetics
  • Oncogene Proteins, Fusion / genetics*
  • RNA, Messenger / genetics*
  • Recurrence
  • Remission Induction
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Translocation, Genetic

Substances

  • Antineoplastic Agents
  • Biomarkers, Tumor
  • DNA Probes
  • MLL-MLLT3 fusion protein, human
  • Oligonucleotides
  • Oncogene Proteins, Fusion
  • RNA, Messenger
  • locked nucleic acid