Annals of Laboratory Medicine

Korean Society for Laboratory Medicine (대한진단검사의학회)
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Development of a Next-generation Sequencing-based Gene Panel Test to Detect Measurable Residual Disease in Acute Myeloid Leukemia

  • Jin Ju Kim (Department of Laboratory Medicine, Yonsei University College of Medicine, Severance Hospital) ;
  • Ji Eun Jang (Division of Hematology, Department of Internal Medicine, Yonsei University College of Medicine, Severance Hospital) ;
  • Hyeon Ah Lee (Department of Laboratory Medicine, Graduate School of Medical Science, Brain Korea 21 PLUS Project, Yonsei University College of Medicine) ;
  • Mi Ri Park (Department of Laboratory Medicine, Yonsei University College of Medicine, Severance Hospital) ;
  • Hye Won Kook (Division of Hematology, Department of Internal Medicine, Yonsei University College of Medicine, Severance Hospital) ;
  • Seung-Tae Lee (Department of Laboratory Medicine, Yonsei University College of Medicine, Severance Hospital) ;
  • Jong Rak Choi (Department of Laboratory Medicine, Yonsei University College of Medicine, Severance Hospital) ;
  • Yoo Hong Min (Division of Hematology, Department of Internal Medicine, Yonsei University College of Medicine, Severance Hospital) ;
  • Saeam Shin (Department of Laboratory Medicine, Yonsei University College of Medicine, Severance Hospital) ;
  • June-Won Cheong (Division of Hematology, Department of Internal Medicine, Yonsei University College of Medicine, Severance Hospital)
  • Received : 2022.08.25
  • Accepted : 2023.01.27
  • Published : 2023.07.01
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Abstract

Background: AML is a heterogeneous disease, and despite intensive therapy, recurrence is still high in AML patients who achieve the criterion for cytomorphologic remission (residual tumor burden [measurable residual disease, MRD]<5%). This study aimed to develop a targeted next-generation sequencing (NGS) panel to detect MRD in AML patients and validate its performance. Methods: We designed an error-corrected, targeted MRD-NGS panel without using physical molecular barcodes, including 24 genes. Fifty-four bone marrow and peripheral blood samples from 23 AML patients were sequenced using the panel. The panel design was validated using reference material, and accuracy was assessed using droplet digital PCR. Results: Dilution tests showed excellent linearity and a strong correlation between expected and observed clonal frequencies (R>0.99). The test reproducibly detected MRD in three dilution series samples, with a sensitivity of 0.25% for single-nucleotide variants. More than half of samples from patients with morphologic remission after one month of chemotherapy had detectable mutations. NGS-MRD positivity for samples collected after one month of chemotherapy tended to be associated with poor overall survival and progression-free survival. Conclusions: Our highly sensitive and accurate NGS-MRD panel can be readily used to monitor most AML patients in clinical practice, including patients without gene rearrangement. In addition, this NGS-MRD panel may allow the detection of newly emerging clones during clinical relapse, leading to more reliable prognoses of AML.

Keywords

Acknowledgement

This study was supported by the Severance Hospital Research Fund for Clinical Excellence (grant number: C-2020-0020) and a Sejin joint research grant of the Department of Laboratory Medicine, Yonsei University College of Medicine (2021-12).

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