title: [Evaluation of methods for detection of NPM1 gene mutations in acute myeloid leukemia].
abstract: The purpose of this study was to establish real-time based methods for detection of NPM1 gene mutation in acute myeloid leukemia (AML). Primers/probes were designed according to the clustered region of NPM1 mutations on exon 12. Two real-time PCR assays, including high resolution melting curve (HRM) and allele-specific PCR (AS-PCR), were developed and clinically evaluated with 89 AML samples, which were parallelly detected by capillary electrophoresis (CE) and sequencing. The results showed that a total of 17 mutation-positive samples were detected, including type A (15 cases), type B (1 case) and type Nm (1 case). HRM assay could detect all mutant types, and the analytical sensitivity was around 5%. In contrast, AS-PCR assay detected only 95% mutant types, but its sensitivity was as high as 0.01%. It is concluded that considering the characteristics of each method as well as the clinical evaluation results, HRM may be used for screening of NPM1 mutations at diagnosis, while the AS-PCR can be used for the MRD quantification during follow-up.
authors.affiliation: School of Life Science, Xiamen University, Xiamen 361005, Fujian Province, China.
authors.forename: Zhi-Peng
authors.initials: ZP
authors.lastname: Li
authors.forename: Xuan
authors.initials: X
authors.lastname: Zhang
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Zhao
authors.forename: Qing-Ge
authors.initials: QG
authors.lastname: Li
authors.forename: Ya-Mei
authors.initials: YM
authors.lastname: Chen
authors.forename: Quan-Yi
authors.initials: QY
authors.lastname: Lu
mesh_terms: D000483:Alleles; D004252:DNA Mutational Analysis; D019075:Electrophoresis, Capillary / Q000379:methods*; D016678:Genome; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis* / Q000235:genetics; D009154:Mutation; D018365:Neoplasm, Residual / Q000175:diagnosis; D009687:Nuclear Proteins / Q000235:genetics*; D000090243:Nucleophosmin; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D000090243:Nucleophosmin