Published online before print July 30, 2009

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Journal of Molecular Diagnostics 2009, Vol. 11, No. 5
Copyright © 2009 American Society for Investigative Pathology & Association for Molecular Pathology
DOI: 10.2353/jmoldx.2009.090043

Rapid Detection of KIT Mutations in Core-Binding Factor Acute Myeloid Leukemia Using High-Resolution Melting Analysis

Óscar Fuster^*, Eva Barragán^*, Pascual Bolufer^*, José Cervera, Maria José Larráyoz, Antonio Jiménez-Velasco, Joaquín Martínez-López^¶, Ana Valencia, Federico Moscardó and Miguel Ángel Sanz

From the Laboratory of Molecular Biology, Department of Medical Pathology, * and the Department of Hematology, Hospital Universitario La Fe, Valencia; the Department of Genetics, Universidad de Navarra, Pamplona; the Department of Hematology, Hospital Universitario Carlos Haya, Málaga; and the Department of Hematology, ^¶ Hospital Universitario 12 de Octubre, Valencia, Spain

The most frequent KIT mutations reported in core-binding factor acute myeloid leukemia are point mutations and insertions/deletions in exons 17 and 8. The vast majority of KIT mutation detection procedures are time-consuming, costly, or with a high lower limit of detection. High-resolution melting (HRM) is a gene scanning method that combines simplicity and rapid identification of genetic variants. We describe an HRM method for the simultaneous screening of exons 8 and 17 KIT mutations and report the results obtained in 69 core-binding factor acute myeloid leukemia patients. Mutation detection was compared with sequencing as the gold standard. The HRM method used high-resolution melting master reagents (Roche) and the LightCycler 480 (Roche) platform. HRM was reproducible, showed a lower limit of detection of 1%, and discriminated all patients with mutated KIT from controls without false positive or false negative results. Additionally, most of the mutations were differentiated from the other mutations. KIT mutations were present in 15.9% of patients, showing a higher incidence in inv(16) (25.8%) than in t(8;21) (7.9%). The presence of a KIT mutation was associated with a high white blood cell count, and adult patients with an exon 17 mutation had a higher incidence of relapse. These findings verify that HRM is a reliable, rapid, and sensitive method for KIT mutation screening. Furthermore, our study corroborates the unfavorable prognosis associated with exon 17 KIT mutations.