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Background:  Molecular genetic alterations with prognostic significance have been described in childhood acute myeloid leukemia (AML). The aim of this study was to establish cost-effective techniques to detect mutations of FMS-like tyrosine kinase 3 ( FLT 3), nucleophosmin 1 ( NPM1 ), and a partial tandem duplication within the mixed-lineage leukemia ( MLL-PTD ) genes in childhood AML.   Procedure:  Ninety-nine children with newly diagnosed AML were included in this study. We developed a fluorescent dye SYTO-82 based high-resolution melting (HRM) curve analysis to detect  FLT 3 internal tandem duplication ( FLT3-ITD ),  FLT 3 tyrosine kinase domain ( FLT3-TKD ), and  NPM1  mutations.  MLL-PTD  was screened by real-time quantitative PCR.   Results:  The HRM methodology correlated well with gold standard Sanger sequencing with less cost. Among the 99 patients studied, the  FLT3-ITD  mutation was associated with significantly worse event-free survival (EFS). Patients with the  NPM1  mutation had significantly better EFS and overall survival. However, HRM was not sensitive enough for minimal residual disease monitoring.   Conclusion:  High-resolution melting was a rapid and efficient method for screening of  FLT 3 and  NPM1  gene mutations. It was both affordable and accurate, especially in resource underprivileged regions. Our results indicated that HRM could be a useful clinical tool for rapid and cost-effective screening of the  FLT3  and  NPM1  mutations in AML patients.

High-Resolution Melting Curve Analysis, a Rapid and Affordable Method for Mutation Analysis in Childhood Acute Myeloid Leukemia