CAT #: FLT3 ITD MRD

FLT3 ITD MRD Testing by NGS

  • Description of Test:

    To track and identify previously detected FLT3 ITD mutations in post treatment follow up samples, a multiplex master mix targeting the juxtamembrane domain of the FLT3 gene is used to amplify DNA extracted from a patient sample.

    Next-generation sequencing of the PCR products is used to identify DNA sequences specific to previously identified mutations detected at diagnosis. Bioinformatics tools facilitate the detection of these specific sequences present at two levels of sensitivity.

    The assay requires a sample taken at diagnosis as well as the post treatment follow up samples. However, in the event of a diagnostic sample not being available, we are still able to detect ITDs.

    • The sensitive assay reliably detects sequences present at 5×10-5.
  • Overview:

    Minimal residual disease (MRD) detection in patients with leukemia has proven to be useful in the clinical management of disease and can facilitate the development of new therapies. Mutations in fms related tyrosine kinase 3 (FLT3) gene are the most common mutations found in acute myeloid leukemia (AML) and are characterized by an aggressive phenotype with a high prevalence of relapse. Internal tandem duplication (ITD) mutations within the juxtamembrane domain are the most common mutations of FLT3. The development of a sensitive and specific assay for FLT3 ITD mutations represents a significant advancement in guiding treatment decisions.

    LabPMM’s FLT3 ITD MRD test is an NGS-based targeted deep sequencing assay that can be used to detect ITDs ranging from 9 bp to >126 bp in size. However, clinical performance of the assay has confirmed ITDs ranging from 3 bp to over 200 bp in size can also in fact be detected. Once a specific ITD (length and sequence) has been identified in a primary sample, it can easily be tracked in subsequent samples at a sensitivity of 10-4 or higher, provided sufficient DNA quantity is tested.

    MRD detection by Next-Generation Sequencing has demonstrated utility in predicting clinical outcomes and in generating clinically actionable results allowing early intervention, confirmation of disease status prior to transplant, and increased confidence in remission status.

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References

  1. The Cancer Genome Atlas Research Network (2013) Genomic and Epigenomic Landscapes of Adult De Novo Acute Myeloid Leukemia. N Engl J Med. 368: 2059–2074.
  2. Konig H. et al. (2015) Targeting FLT3 to treat leukemia. Expert Opin Ther Targets 19:37-54.

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