{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,19]],"date-time":"2026-10-19T03:39:55Z","timestamp":1792381195572,"version":"4.4.0"},"reference-count":26,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2026,6,15]],"date-time":"2026-06-15T00:00:00Z","timestamp":1781481600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,6,15]],"date-time":"2026-06-15T00:00:00Z","timestamp":1781481600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Int J Lab Hematology"],"published-print":{"date-parts":[[2026,10]]},"abstract":"ABSTRACT<\/jats:title>\n \n Introduction<\/jats:title>\n \n Quantification and monitoring of\n NPM1<\/jats:italic>\n mutations represent a key tool in the management of acute myeloid leukemia (AML), offering strong prognostic value and serving as a marker of minimal residual disease (MRD). Digital PCR (dPCR) provides absolute quantification with high sensitivity and specificity, offering advantages over conventional qPCR\u2010based methods, particularly for rare or atypical\n NPM1<\/jats:italic>\n mutations.\n <\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n We adapted and evaluated a digital PCR strategy including both monoplex and multiplex assays targeting the most common\n NPM1<\/jats:italic>\n mutations (types A, B, D) as well as rare variants. A touch\u2010down PCR protocol was optimized to harmonize the dPCR protocol and enhance specificity. Assay performance was evaluated in terms of limit of detection (LoD), limit of quantification (LoQ), linearity, and specificity, and compared to quantitative PCR (qPCR).\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n The multiplex dPCR assay allowed detection of all clinically relevant\n NPM1<\/jats:italic>\n mutations with an LoD of 0.01%, while monoplex type A, B and D assays reached 0.001% for type A, B and D. The method showed strong linearity (\n R<\/jats:italic>\n 2<\/jats:sup>\n \u2009>\u20090.99 for all targets) and high concordance with qPCR (R\n 2<\/jats:sup>\n \u2009=\u20090.9967). In patient samples, dPCR demonstrated improved sensitivity over qPCR, detecting low\u2010level NPM1 mutations in samples previously negative by qPCR, and enabled early relapse detection.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n \n The approach enabled early relapse prediction in selected patients and improved molecular follow\u2010up, even for rare\n NPM1<\/jats:italic>\n mutations not covered by standard qPCR panels. It simplifies laboratory routine and improves MRD assessment, according to current ELN guidelines and the growing need for personalized molecular monitoring in AML.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1111\/ijlh.70161","type":"journal-article","created":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T05:50:11Z","timestamp":1781589011000},"page":"1149-1157","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Optimization of Analytical Performances and Routine Laboratory Implementation of\n \n NPM1<\/i>\n <\/scp>\n Mutation Detection by Digital\n PCR<\/scp>\n in the Diagnosis and Monitoring of\n \n NPM1<\/i>\n <\/scp>\n \u2010Mutated Acute Myeloid Leukemias"],"prefix":"10.1111","volume":"48","author":[{"given":"Chlo\u00e9","family":"Ferrari","sequence":"first","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"},{"name":"Universit\u00e9 de Caen, Normandie Universit\u00e9 Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brice","family":"Coulleray","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pauline","family":"Kerneves","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carole","family":"Geara","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dina","family":"Naguib","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gu\u00e9naelle","family":"Levallet","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"},{"name":"Universit\u00e9 de Caen, Normandie Universit\u00e9 Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth","family":"Battersby","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"},{"name":"Universit\u00e9 de Caen, Normandie Universit\u00e9 Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sylvain","family":"Chantepie","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie, Institut D'h\u00e9matologie Bas Normand (IHBN) Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yohann","family":"Repess\u00e9","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"},{"name":"Universit\u00e9 de Caen, Normandie Universit\u00e9 Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Coralie","family":"Nectoux","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5967-8225","authenticated-orcid":false,"given":"Elsa","family":"Maitre","sequence":"additional","affiliation":[{"name":"Centre Hospitalier Universitaire de Caen Normandie Caen France"},{"name":"Universit\u00e9 de Caen, Normandie Universit\u00e9 Caen France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,6,15]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.5858\/arpa.2021\u20100601\u2010OA"},{"issue":"1","key":"e_1_2_11_3_1","first-page":"28","article-title":"Prognostic Relevance of NPM1, CEBPA, and FLT3 Mutations in Cytogenetically Normal Adult AML Patients","volume":"13","author":"Ningombam 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