Related Experiment Video
Updated: Jul 10, 2025
Next Generation Sequencing for the Detection of Actionable Mutations in Solid and Liquid Tumors
Published on: September 20, 2016
Standardization of Molecular MRD Levels in AML Using an Integral Vector Bearing ABL and the Mutation of Interest
Boaz Nachmias1, Svetlana Krichevsky1, Moshe E Gatt1
1Department of Hematology, Hadassah Medical Center and Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem 91200, Israel.
Abstract:
Quantitative PCR for specific mutation is being increasingly used in Acute Myeloid Leukemia (AML) to assess Measurable Residual Disease (MRD), allowing for more tailored clinical decisions. To date, standardized molecular MRD is limited to typical NPM1 mutations and core binding factor translocations, with clear prognostic and clinical implications. The monitoring of other identified mutations lacks standardization, limiting its use and incorporation in clinical trials. To overcome this problem, we designed a plasmid bearing both the sequence of the mutation of interest and the ABL reference gene. This allows the use of commercial standards for ABL to determine the MRD response in copy number. We provide technical aspects of this approach as well as our experience with 19 patients with atypical NPM1, RUNX1 and IDH1/2 mutations. In all cases, we demonstrate a correlation between response and copy number. We further demonstrate how copy number monitoring can modulate the clinical management. Taken together, we provide proof of concept of a novel yet simple tool, which allows in-house MRD monitoring for identified mutations, with ABL-based commercial standards. This approach would facilitate large multi-center studies assessing the clinical relevance of selected MRD monitoring.
Insights
A novel plasmid-based quantitative PCR method enables in-house monitoring of Measurable Residual Disease (MRD) in Acute Myeloid Leukemia (AML) for various mutations. This tool standardizes MRD assessment, aiding clinical decisions and multi-center trials.
Area of Science:
- Hematology
- Molecular Biology
- Oncology
Background:
- Quantitative PCR is crucial for assessing Measurable Residual Disease (MRD) in Acute Myeloid Leukemia (AML), guiding clinical decisions.
- Standardized MRD monitoring is currently limited to specific mutations (NPM1, CBF), hindering the assessment of other mutations.
Purpose of the Study:
- To develop and validate a novel, standardized method for in-house MRD monitoring in AML patients with diverse mutations.
- To establish a reliable tool for assessing MRD response using ABL reference gene and commercial standards.
Main Methods:
- A plasmid construct containing both the mutation of interest and the ABL reference gene was designed.
- Quantitative PCR was performed using commercial ABL standards to determine MRD response in copy number.
- The method was applied to 19 AML patients with atypical NPM1, RUNX1, and IDH1/2 mutations.
Main Results:
- The developed method demonstrated a correlation between MRD response and copy number in all tested patients.
- Copy number monitoring was shown to influence clinical management decisions.
- The approach provides a standardized, in-house solution for MRD monitoring of identified mutations.
Conclusions:
- This plasmid-based quantitative PCR approach offers a simple, novel tool for in-house MRD monitoring in AML.
- The method facilitates standardized assessment of MRD for a broader range of mutations, supporting clinical trials.
- This tool can improve clinical decision-making and advance research into the prognostic significance of MRD in AML.
More Related Videos
09:57Comprehensive Protocol to Sample and Process Bone Marrow for Measuring Measurable Residual Disease and Leukemic Stem Cells in Acute Myeloid Leukemia
Published on: March 5, 2018
08:46Genetic Variant Detection in the CALR gene using High Resolution Melting Analysis
Published on: August 26, 2020
