Feasible and Rapid Screening of and Mutations by High-Resolution Melting for Patients with Acute Myeloid Leukemia.

: In recent years, numerous recurrently mutated genes have been identified in acute myeloid leukemia (AML), some of which, such as and , serve as therapeutic targets, offering new treatment options. Rapid mutational analysis is crucial for timely and optimal therapy selection. This study aims to develop and validate a rapid, cost-effective, and sensitive screening method for detecting , , and -TKD2 mutations using polymerase chain reaction (PCR) and high-resolution melting curve analysis (HRM). : A PCR-HRM assay was developed to simultaneously detect mutations in , , and -TKD2. The method was applied to a cohort of 1363 AML patients, and its performance, including turnaround time, was evaluated through comparison with next-generation sequencing (NGS) results. : The PCR-HRM method demonstrated a positive percent agreement of 98%, 98%, and 92% for , , and , respectively, and a negative percent agreement of 100% for all three genes compared to NGS. No false positives were observed, and false negatives were detected in less than 1% of cases, mostly in -TKD2, all occurring below the established limit of detection. The turnaround time and cost of PCR-HRM were significantly lower than those of NGS. : This method offers a highly sensitive, specific, and time-efficient approach for the simultaneous detection of , , and -TKD2 mutations in AML patients. Its rapid turnaround time and cost-effectiveness make it a valuable tool for routine clinical screening, facilitating timely and targeted treatment decisions.