Rapid Detection of N-RAS Gene Common Mutations in Acute Myeloid Leukemia (AML) Using High Resolution Melting (HRM) Method

Abstract

Objective: Acute myeloid leukemia is caused by the clonal proliferation of undifferentiated myeloid hematopoietic precursors. AML prognosis is highly involved in the treatment response and is determined by mutations in several genessuch as N-RAS. This study aims to identify the distribution of common N-RAS mutations (codons 12, 13, and 61) inAML patients using the HRM method and confirm this method’s efficiency for mutation detection by comparing itsresults with the sequencing data as the Gold standard method. Methods: Peripheral blood samples were taken from 50newly diagnosed AML patients. Mononuclear cells were isolated from samples, and DNA was extracted. Then, mutationdetection was investigated using the HRM method. Efficacy of the HRM method in mutation detection was determinedin comparison with direct sequencing. Results: N-RAS mutations were detected in 7 of the 50 samples (14%). Mostof the mutations were found in codon 12 (57.14%), and 28.57% and 14.28% of mutations were in codons 61 and 13,respectively. There was no statistically significant association between patients’ demographic data and HRM results.Conclusion: According to mutation detection results and the HRM results confirmation with the sequencing method,this method can be introduced as an efficient, low-cost, and fast method for detecting common mutations