AUTHOR=Liu Yin , Tang Jingyan , Wakamatsu Peter , Xue Huiliang , Chen Jing , Gaynon Paul S. , Shen Shuhong , Sun Weili 

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

JOURNAL=Frontiers in Pediatrics

VOLUME=2

YEAR=2014

URL=https://www.frontiersin.org/journals/pediatrics/articles/10.3389/fped.2014.00096

DOI=10.3389/fped.2014.00096

ISSN=2296-2360

ABSTRACT=<p><bold>Background:</bold> 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 (<italic>FLT</italic>3), nucleophosmin 1 (<italic>NPM1</italic>), and a partial tandem duplication within the mixed-lineage leukemia (<italic>MLL-PTD</italic>) genes in childhood AML.</p><p><bold>Procedure:</bold> 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 <italic>FLT</italic>3 internal tandem duplication (<italic>FLT3-ITD</italic>), <italic>FLT</italic>3 tyrosine kinase domain (<italic>FLT3-TKD</italic>), and <italic>NPM1</italic> mutations. <italic>MLL-PTD</italic> was screened by real-time quantitative PCR.</p><p><bold>Results:</bold> The HRM methodology correlated well with gold standard Sanger sequencing with less cost. Among the 99 patients studied, the <italic>FLT3-ITD</italic> mutation was associated with significantly worse event-free survival (EFS). Patients with the <italic>NPM1</italic> mutation had significantly better EFS and overall survival. However, HRM was not sensitive enough for minimal residual disease monitoring.</p><p><bold>Conclusion:</bold> High-resolution melting was a rapid and efficient method for screening of <italic>FLT</italic>3 and <italic>NPM1</italic> 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 <italic>FLT3</italic> and <italic>NPM1</italic> mutations in AML patients.</p>