title: Fragment length analysis screening for detection of CEBPA mutations in intermediate-risk karyotype acute myeloid leukemia.
abstract: During last years, molecular markers have been increased as prognostic factors routinely screened in acute myeloid leukemia (AML). Recently, an increasing interest has been reported in introducing to clinical practice screening for mutations in the CCAAT/enhancer-binding protein α (CEBPA) gene in AML, as it seems to be a good prognostic factor. However, there is no reliable established method for assessing CEBPA mutations during the diagnostic work-up of AMLs. We describe here a straightforward and reliable fragment analysis method based in PCR capillary electrophoresis (PCR-CE) for screening of CEBPA mutations; moreover, we present the results obtained in 151 intermediate-risk karyotype AML patients (aged 16-80 years). The method gave a specificity of 100% and sensitivity of 93% with a lower detection limit of 1-5% for CEBPA mutations. The series found 19 mutations and four polymorphisms in 12 patients, seven of whom (58%) presented two mutations. The overall frequency of CEBPA mutations in AML was 8% (n = 12). CEBPA mutations showed no coincidence with FLT3-ITD or NPM1 mutations. CEBPA mutation predicted better disease-free survival in the group of patients without FLT3-ITD, NPM, or both genes mutated (HR 3.6, IC 95%; 1.0-13.2, p = 0.05) and better overall survival in patients younger than 65 of this group without molecular markers (HR 4.0, IC 95%; 1.0-17.4, p = 0.05). In conclusion, the fragment analysis method based in PCR-CE is a rapid, specific, and sensitive method for CEBPA mutation screening and our results confirm that CEBPA mutations can identify a subgroup of patients with favorable prognosis in AML with intermediate-risk karyotype.
authors.affiliation: Laboratory of Molecular Biology, Department of Medical Pathology, Hospital Universitario La Fe, Escuela de enfermería 7ª planta., Avd. Campanar 21, Valencia 46009, Spain.
authors.forename: Oscar
authors.initials: O
authors.lastname: Fuster
authors.forename: Eva
authors.initials: E
authors.lastname: Barragán
authors.forename: Pascual
authors.initials: P
authors.lastname: Bolufer
authors.forename: Esperanza
authors.initials: E
authors.lastname: Such
authors.forename: Ana
authors.initials: A
authors.lastname: Valencia
authors.forename: Mariam
authors.initials: M
authors.lastname: Ibáñez
authors.forename: Sandra
authors.initials: S
authors.lastname: Dolz
authors.forename: Inmaculada
authors.initials: I
authors.lastname: de Juan
authors.forename: Antonio
authors.initials: A
authors.lastname: Jiménez
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Gómez
authors.forename: Ismael
authors.initials: I
authors.lastname: Buño
authors.forename: Joaquín
authors.initials: J
authors.lastname: Martínez
authors.forename: José
authors.initials: J
authors.lastname: Cervera
authors.forename: Pau
authors.initials: P
authors.lastname: Montesinos
authors.forename: Federico
authors.initials: F
authors.lastname: Moscardó
authors.forename: Miguel Ángel
authors.initials: MÁ
authors.lastname: Sanz
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics*; D018572:Disease-Free Survival; D019075:Electrophoresis, Capillary / Q000379:methods; D005260:Female; D005819:Genetic Markers; D006801:Humans; D059785:Karyotype; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D000090243:Nucleophosmin; D010446:Peptide Fragments / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D011379:Prognosis; D017422:Sequence Analysis, DNA / Q000379:methods*; D015996:Survival Rate; D055815:Young Adult
chemical_list: D022763:CCAAT-Enhancer-Binding Protein-alpha; D005819:Genetic Markers; C000716967:NPM1 protein, human; D010446:Peptide Fragments; D000090243:Nucleophosmin