title: Detection of nucleophosmin 1 mutations by quantitative real-time polymerase chain reaction versus capillary electrophoresis: a comparative study.
abstract: BACKGROUND
  Nucleophosmin 1 (NPM1) is the most commonly mutated gene in acute myeloid leukemia. Detection of NPM1 mutations is useful for stratifying patients for therapy, predicting prognosis, and assessing for minimal residual disease. Several methods have been developed to rapidly detect NPM1 mutations in genomic DNA and/or messenger RNA specimens.
  
  
  OBJECTIVE
  To directly compare a quantitative real-time polymerase chain reaction (qPCR) assay with a widely used capillary electrophoresis assay for detecting NPM1 mutations.
  
  
  METHODS
  We adopted and modified a qPCR assay designed to detect the 6 most common NPM1 mutations and performed the assay in parallel with capillary electrophoresis assay in 207 bone marrow aspirate or peripheral blood samples from patients with a range of hematolymphoid neoplasms.
  
  
  RESULTS
  The qPCR assay demonstrated a higher analytical sensitivity than the capillary electrophoresis 1/1000 versus 1/40, respectively. The capillary electrophoresis assay generated 10 equivocal results that needed to be repeated, whereas the qPCR assay generated only 1 equivocal result. After test conditions were optimized, the qPCR and capillary electrophoresis methods produced 100% concordant results, 85 positive and 122 negative.
  
  
  CONCLUSIONS
  Given the higher analytical sensitivity and specificity of the qPCR assay, that assay is less likely to generate equivocal results than the capillary electrophoresis assay. Moreover, the qPCR assay is quantitative, faster, cheaper, less prone to contamination, and well suited for monitoring minimal residual disease.
authors.affiliation: Department of Hematopathology, The University of Texas M. D. Anderson Cancer Center, Houston, 77054, USA.
authors.forename: Fareed H
authors.initials: FH
authors.lastname: Barakat
authors.forename: Rajyalakshmi
authors.initials: R
authors.lastname: Luthra
authors.forename: C Cameron
authors.initials: CC
authors.lastname: Yin
authors.forename: Bedia A
authors.initials: BA
authors.lastname: Barkoh
authors.forename: Seema
authors.initials: S
authors.lastname: Hai
authors.forename: Waqar
authors.initials: W
authors.lastname: Jamil
authors.forename: Yaminiben I
authors.initials: YI
authors.lastname: Bhakta
authors.forename: Su
authors.initials: S
authors.lastname: Chen
authors.forename: L Jeffrey
authors.initials: LJ
authors.lastname: Medeiros
authors.forename: Zhuang
authors.initials: Z
authors.lastname: Zuo
mesh_terms: D001854:Bone Marrow Cells / Q000737:chemistry / Q000473:pathology; D017931:DNA Primers; D015342:DNA Probes; D004273:DNA, Neoplasm / Q000032:analysis; D019075:Electrophoresis, Capillary / Q000379:methods*; D019337:Hematologic Neoplasms / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D009154:Mutation; D009687:Nuclear Proteins / Q000097:blood / Q000235:genetics*; D000090243:Nucleophosmin; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D004273:DNA, Neoplasm; D009687:Nuclear Proteins; D000090243:Nucleophosmin