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A Novel Tandem Duplication Assay to Detect Minimal Residual Disease in FLT3/ITD AML

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Abstract

Background

Internal tandem duplication (ITD) of the fms-related tyrosine kinase 3 (FLT3) gene is associated with a poor prognosis in acute myeloid leukemia (AML) patients with a normal karyotype. The current standard polymerase chain reaction (PCR) assay for FLT3/ITD detection is not sufficiently sensitive to monitor minimal residual disease (MRD). Clone-specific assays may have sufficient sensitivity but are not practical to implement, since each clone-specific primer/probe requires clinical validation.

Objective

To develop an assay for clinical molecular diagnostics laboratories to monitor MRD in FLT3/ITD AMLs.

Methods

We designed a simple novel assay, tandem duplication PCR (TD-PCR), and tested its sensitivity, specificity, and clinical utility in FLT3/ITD AML patients.

Results

TD-PCR was capable of detecting a single ITD molecule and was applicable to 75 % of ITD mutants tested. TD-PCR detected MRD in bone marrow prior to patient relapse. TD-PCR also identified low-level ITD mutants not only in FLT3/ITD AMLs but also in initial diagnostic specimens that were reportedly negative by the standard assay in patients who progressed with the same ITDs detected by the TD-PCR assay.

Conclusion

Detection of MRD by TD-PCR may guide patient selection for early clinical intervention. In contrast to clone-specific approaches, the TD-PCR assay can be more easily validated for MRD detection in clinical laboratories because it uses standardized primers and a universal positive control. In addition, our findings on multi-clonality and low-level ITDs suggest that further studies are warranted to elucidate their clinical/biological significance.

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Author notes

    Authors and Affiliations

    1. Division of Molecular Pathology, Department of Pathology, Johns Hopkins University School of Medicine, Park SB202, 600 North Wolfe Street, Baltimore, MD, 21287, USA

      Ming-Tseh Lin, Li-Hui Tseng, Jonathan C. Dudley, Stacey Riel, Harrison Tsai, Gang Zheng & Christopher D. Gocke

    2. Department of Medical Genetics, National Taiwan University Hospital, Taipei, Taiwan

      Li-Hui Tseng

    3. Department of Pathology, Massachusetts General Hospital, Boston, MA, USA

      Jonathan C. Dudley

    4. Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

      Keith W. Pratz, Mark J. Levis & Christopher D. Gocke

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    1. Ming-Tseh Lin
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    Correspondence to Christopher D. Gocke.

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    Funding

    The authors M-TL and CDG gratefully acknowledge funding received from the US National Institutes of Health (Grant Nos. R21HG004315 and R21HG005745 to CDG) and the National Cancer Institute at the National Institutes of Health (Grant No. 1UM1CA186691-01 to M-TL and CDG).

    Conflict of interest

    All authors (M-TL, L-HT, JCD, SR, HT, GZ, KWP, MJL, and CDG) report that they have no conflicts of interest that are directly relevant to the content of this article.

    Ethical approval and informed consent

    The Johns Hopkins Medicine institutional review board granted approval for this study.

    Additional information

    M.-T. Lin and L.-H. Tseng contributed equally to this article.

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    Lin, MT., Tseng, LH., Dudley, J.C. et al. A Novel Tandem Duplication Assay to Detect Minimal Residual Disease in FLT3/ITD AML. Mol Diagn Ther 19, 409–417 (2015). https://doi.org/10.1007/s40291-015-0170-3

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    • DOI: https://doi.org/10.1007/s40291-015-0170-3

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