Molecular Genetic Tests for FLT3, NPM1, and CEBPA in Acute Myeloid Leukemia

  • Qing Zhang
  • Shaochun Bai
  • Gail H. Vance
Part of the Methods in Molecular Biology book series (MIMB, volume 999)


Patients with acute myeloid leukemia (AML) and a normal karyotype constitute the single largest cytogenetic group of AML. It is important to identify prognostic markers that predict patients’ outcome more precisely. The presence of mutations in FLT3 (FMS-like tyrosine kinase 3), NPM1 (Nucleophosmin), and CEBPA (CCAAT/enhancer-binding protein alpha) genes hold prognostic significance in patients with AML and normal cytogenetics. Therefore, mutation identification may help to optimize therapeutic approaches in this group of patients. Polymerase chain reaction (PCR)-based fragment length analysis for mutations in FLT3 and NPM1 has been shown to be a fast and sensitive method, while nucleotide sequencing represents a gold standard for CEBPA heterogeneous mutational screening. We describe both fragment length assay and sequencing methods for mutational analysis of these three genes.

Key words

Acute myeloid leukemia (AML) FLT3 NPM1 CEBPA PCR Capillary electrophoresis DNA sequencing Mutational detection 



The authors wish to thank Dr. Stephen Dlouhy for his review of the manuscript.


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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Qing Zhang
    • 1
  • Shaochun Bai
    • 1
  • Gail H. Vance
    • 1
  1. 1.Department of Medical and Molecular GeneticsIndiana University School of MedicineIndianapolisUSA

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