Ultrasensitive quantitation of FLT3-ITD mutation in patients with acute myeloid leukemia using ddPCR

Mol Biol Rep. 2023 Jul;50(7):6097-6105. doi: 10.1007/s11033-023-08534-x. Epub 2023 Jun 10.

Abstract

Background: FLT3-ITD mutations occur in 45-50% of cytogenetically normal AML patients. Conventional fragment analysis using capillary electrophoresis is routinely used to quantitate FLT3-ITD mutations. Fragment analysis however has limited sensitivity.

Methods and results: Here, FLT3-ITD was quantified in AML patients using an in-house developed ultra-sensitive droplet digital polymerase chain reaction assay (ddPCR). The allelic ratio of FLT3-ITD was also absolutely measured by both Fragment analysis and ddPCR. The sensitivity of ddPCR in quantitation of FLT3-ITD mutation was superior to Fragment analysis.

Conclusion: This study demonstrates the feasibility of using the described in-house ddPCR method to quantify the FLT3-ITD mutation and measure FLT3-ITD AR in AML patients.

Keywords: AML; FLT3-ITD; Fragment analysis; ddPCR.

MeSH terms

  • Humans
  • Leukemia, Myeloid, Acute* / genetics
  • Mutation / genetics
  • Nuclear Proteins* / genetics
  • Nucleophosmin
  • Polymerase Chain Reaction
  • fms-Like Tyrosine Kinase 3 / genetics

Substances

  • Nuclear Proteins
  • Nucleophosmin
  • fms-Like Tyrosine Kinase 3
  • FLT3 protein, human