{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T13:25:17Z","timestamp":1770297917030,"version":"3.49.0"},"reference-count":29,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T00:00:00Z","timestamp":1747180800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Scientific Foundation of the Spanish Association Against Cancer","award":["PRDVA246268GIL"],"award-info":[{"award-number":["PRDVA246268GIL"]}]},{"name":"Scientific Foundation of the Spanish Association Against Cancer","award":["PMP22\/00069"],"award-info":[{"award-number":["PMP22\/00069"]}]},{"name":"Instituto de Salud Carlos III, Spain","award":["PRDVA246268GIL"],"award-info":[{"award-number":["PRDVA246268GIL"]}]},{"name":"Instituto de Salud Carlos III, Spain","award":["PMP22\/00069"],"award-info":[{"award-number":["PMP22\/00069"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background: In recent years, numerous recurrently mutated genes have been identified in acute myeloid leukemia (AML), some of which, such as FLT3 and IDH1\/2, 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 IDH1, IDH2, and FLT3-TKD2 mutations using polymerase chain reaction (PCR) and high-resolution melting curve analysis (HRM). Methods: A PCR-HRM assay was developed to simultaneously detect mutations in IDH1, IDH2, and FLT3-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. Results: The PCR-HRM method demonstrated a positive percent agreement of 98%, 98%, and 92% for IDH1, IDH2, and FLT3-TKD2, 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 FLT3-TKD2, all occurring below the established limit of detection. The turnaround time and cost of PCR-HRM were significantly lower than those of NGS. Conclusions: This method offers a highly sensitive, specific, and time-efficient approach for the simultaneous detection of IDH1, IDH2, and FLT3-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.<\/jats:p>","DOI":"10.3390\/diagnostics15101230","type":"journal-article","created":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T03:59:29Z","timestamp":1747195169000},"page":"1230","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Feasible and Rapid Screening of IDH1\/2 and FLT3-TKD2 Mutations by High-Resolution Melting for Patients with Acute Myeloid Leukemia"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2097-4796","authenticated-orcid":false,"given":"Jos\u00e9 Vicente","family":"Gil","sequence":"first","affiliation":[{"name":"Accredited Research Group on Hematology, Instituto de Investigaci\u00f3n Sanitaria la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sandra","family":"de las Heras","sequence":"additional","affiliation":[{"name":"Accredited Research Group on Hematology, Instituto de Investigaci\u00f3n Sanitaria la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alberto","family":"Miralles","sequence":"additional","affiliation":[{"name":"Accredited Research Group on Hematology, Instituto de Investigaci\u00f3n Sanitaria la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8144-7995","authenticated-orcid":false,"given":"Claudia","family":"Sargas","sequence":"additional","affiliation":[{"name":"Accredited Research Group on Hematology, Instituto de Investigaci\u00f3n Sanitaria la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0106-7296","authenticated-orcid":false,"given":"Marta","family":"Llop","sequence":"additional","affiliation":[{"name":"Molecular Biology Unit, Clinical Analysis Service, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red de C\u00e1ncer, CIBERONC Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2139-8540","authenticated-orcid":false,"given":"Rebeca","family":"Rodr\u00edguez-Veiga","sequence":"additional","affiliation":[{"name":"Hematology Department, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-6930-8059","authenticated-orcid":false,"given":"Laura","family":"Torres-Mi\u00f1ana","sequence":"additional","affiliation":[{"name":"Hematology Department, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Blanca","family":"Boluda","sequence":"additional","affiliation":[{"name":"Hematology Department, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Isabel","family":"Cano-Ferri","sequence":"additional","affiliation":[{"name":"Hematology Department, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2869-369X","authenticated-orcid":false,"given":"Evelyn","family":"Acu\u00f1a-Cruz","sequence":"additional","affiliation":[{"name":"Hematology Department, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Irene","family":"Navarro","sequence":"additional","affiliation":[{"name":"Hematology Department, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pilar","family":"Lloret-Madrid","sequence":"additional","affiliation":[{"name":"Hematology Department, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pau","family":"Montesinos","sequence":"additional","affiliation":[{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red de C\u00e1ncer, CIBERONC Instituto de Salud Carlos III, 28029 Madrid, Spain"},{"name":"Hematology Department, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eva","family":"Barrag\u00e1n","sequence":"additional","affiliation":[{"name":"Accredited Research Group on Hematology, Instituto de Investigaci\u00f3n Sanitaria la Fe, 46026 Valencia, Spain"},{"name":"Molecular Biology Unit, Clinical Analysis Service, Hospital Universitario y Polit\u00e9cnico la Fe, 46026 Valencia, Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red de C\u00e1ncer, CIBERONC Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1158\/2159-8290.CD-19-1011","article-title":"Advances in the Treatment of Acute Myeloid 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