title: High-resolution melting curve analysis, a rapid and affordable method for mutation analysis in childhood acute myeloid leukemia.
abstract: 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 (FLT3), nucleophosmin 1 (NPM1), and a partial tandem duplication within the mixed-lineage leukemia (MLL-PTD) genes in childhood AML.
  
  
  METHODS
  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 FLT3 internal tandem duplication (FLT3-ITD), FLT3 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.
  
  
  CONCLUSIONS
  High-resolution melting was a rapid and efficient method for screening of FLT3 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.
authors.affiliation: Department of Hematology/Oncology, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine , Shanghai , China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Hematology/Oncology, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine , Shanghai , China.
authors.forename: Jingyan
authors.initials: J
authors.lastname: Tang
authors.affiliation: Division of Hematology, Oncology and Blood and Marrow Transplantation, Children's Center for Cancer and Blood Disease, Children's Hospital Los Angeles, Keck School of Medicine, University of Southern California , Los Angeles, CA , USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Wakamatsu
authors.affiliation: Department of Hematology/Oncology, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine , Shanghai , China.
authors.forename: Huiliang
authors.initials: H
authors.lastname: Xue
authors.affiliation: Department of Hematology/Oncology, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine , Shanghai , China.
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.affiliation: Division of Hematology, Oncology and Blood and Marrow Transplantation, Children's Center for Cancer and Blood Disease, Children's Hospital Los Angeles, Keck School of Medicine, University of Southern California , Los Angeles, CA , USA.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Gaynon
authors.affiliation: Department of Hematology/Oncology, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine , Shanghai , China.
authors.forename: Shuhong
authors.initials: S
authors.lastname: Shen
authors.affiliation: Division of Hematology, Oncology and Blood and Marrow Transplantation, Children's Center for Cancer and Blood Disease, Children's Hospital Los Angeles, Keck School of Medicine, University of Southern California , Los Angeles, CA , USA.
authors.forename: Weili
authors.initials: W
authors.lastname: Sun
keywords: FLT3; MLL; NPM1; acute myeloid leukemia; childhood; high-resolution melting curve analysis