title: [Establishment of a rapid and easy method for simultaneous detection of FLT3-ITD and NPM1 gene mutations in acute myeloid leukemia].
abstract: OBJECTIVE
  To establish a stable, rapid multiplex PCR assay combined with PAGE gel electrophoresis for simultaneously detecting FLT3-ITD and NPM1 mutations in acute myeloid leukemia (AML).
  
  
  METHODS
  Capillary electrophoresis (CE) and PAGE gel electrophoresis were simultaneously used to analyze  FLT3-ITD and NPM1 mutations in 117 de novo AML patients with normal cytogenetic findings.
  
  
  RESULTS
  For certain mutations, the length of mutated double-stranded DNA is longer than wild-type DNA. Since FLT3-mut (420 bp) is longer than FLT3-wt (327-332 bp), and  NPM1-mut (172 bp) is longer than NPM1-wt (168 bp), heteroduplex will move more slowly during PAGE gel electrophoresis than homoduplex. Therefore the mutations may be detected. A total of 117 CN-AML patients were analyzed with CE and PAGE gel electrophoresis, and the results were identical, which included 18 (15.4%) patients with FLT3-ITD+/NPM1-, 19 (16.2%) patients with  FLT3-ITD+/NPM1+, 25 (21.4%) patients with FLT3-ITD-/NPM1+, and 55 (47.0%) patients with FLT3-ITD-/NPM1-.
  
  
  CONCLUSIONS
  Both types of electrophoresis assays may provide a rapid and handy assay for simultaneous detection of FLT3-ITD and NPM1 mutations. CE is relatively sensitive, stable; while PAGE electrophoresis is relatively simple, cheap, and reliable, which may be suitable for primary hospitals and preliminary screening.
authors.affiliation: Department of Hematology, Ningbo first hospital, Ningbo, Zhejiang, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Lu
authors.forename: Qiong
authors.initials: Q
authors.lastname: Wang
authors.forename: Qi-tian
authors.initials: QT
authors.lastname: Mu
authors.forename: Meng-xia
authors.initials: MX
authors.lastname: Yu
authors.forename: Qin
authors.initials: Q
authors.lastname: Huang
authors.forename: Jie
authors.initials: J
authors.lastname: Jin
mesh_terms: D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005260:Female; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics*; D000090243:Nucleophosmin; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics*
chemical_list: C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D000090243:Nucleophosmin; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3