Evidence map›Paper›PMID 42545548›Full record

ArticleMolecular biology reports2026

Analytical validation of a modified wild-type blocker qpcr assay for the sensitive DNA-based detection of NPM1 type A in acute myeloid leukemia.

Pejman Hamedi-Asl, Dariush Hamedi-Asl, Shaharbano Rostami, Mahmood Barati, Ali Amini, Davod Jafari, Fatemeh Damerchiloo, Rima Manafi, Majid Safa

Abstract readValidation Study
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In one paragraph

Article in Molecular biology reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

9 authors.

Pejman Hamedi-AslDepartment of Hematology and Blood Banking, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran.
Dariush Hamedi-AslDepartment of Pathology and Molecular Medicine at Behsotun lab, Karaj, Iran.
Shaharbano RostamiHematology, Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Mahmood BaratiDepartment of Medical Biotechnology, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran.
Ali AminiDepartment of Hematology and Blood Banking, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran.
Davod JafariOncopathology research center, Iran University of Medical Sciences, Tehran, Iran.
Fatemeh DamerchilooDepartment of Hematology and Blood Banking, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran.
Rima ManafiDepartment of Hematology and Blood Banking, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran.
Majid SafaDepartment of Hematology and Blood Banking, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran. majidsafa@gmail.com.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundThis study describes the development and analytical validation of a DNA-based quantitative PCR (qPCR) assay for detecting the NPM1 Type A mutation (NPM1-A METHODS AND

resultsThe modified WTB was synthesized as a 3'-ddC-terminated LNA/DNA chimera and used together with a 6-FAM-MGB-Eclipse hydrolysis probe. The analytical performance of the WTB-qPCR assay was compared with that of an otherwise identical qPCR assay lacking WTB. Key parameters such as linearity, precision, limit of detection (LoD), and limit of quantification (LoQ) were evaluated following the guidelines outlined in CLSI EP17-A2. Concurrently, modified WTB was implemented in Sanger sequencing to improve the sensitivity of mutation detection. Incorporation of WTB yielded an LoD of 0.00302% and an LoQ of 0.00528%, corresponding to approximately 3.02 and 5.28 NPM1-A

conclusionThe modified WTB-qPCR assay offers a rapid, highly sensitive, and cost-effective DNA-based method for MRD monitoring in NPM1-A-mutated AML. Its strong analytical performance and straightforward workflow support its implementation as a routine diagnostic tool in clinical laboratories.

Indexed as

Leukemia, Myeloid, AcuteNuclear ProteinsReal-Time Polymerase Chain ReactionAllelesDNA Mutational AnalysisHumansLimit of DetectionMutationNucleophosminReproducibility of ResultsSensitivity and SpecificityNPM1 protein, humanNuclear ProteinsNucleophosminLimit of detectionMeasurable residual diseaseModified wild-type blockerNPM1-AQuantitative PCRSanger sequencing

Identifiers

PMID42545548

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