title: Detection of BRAF V600E mutation by pyrosequencing.
abstract: BACKGROUND
  Detection of the V600E hotspot mutation in BRAF oncogene is extremely useful for the screening of hereditary non-polyposis colorectal cancer (Lynch's syndrome) and for the prediction of sensitivity to MEK inhibitors. Here we describe a method for detecting this mutation based upon pyrosequencing technology.
  
  
  METHODS
  The efficiency of pyrosequencing for detecting BRAF V600E mutations was compared with the conventional dideoxy sequencing method in 12 tumour cell lines and in 108 colorectal tumours.
  
  
  RESULTS
  The results from pyrosequencing were 100% concordant with those from dideoxy sequencing. This method was capable of detecting BRAF V600E mutations at a much lower ratio of mutant to wild-type alleles (1:50) than dideoxy sequencing (1:5) while being considerably faster and less expensive.
  
  
  CONCLUSIONS
  Pyrosequencing offers a specific, sensitive, rapid and cost-effective alternative to dideoxy sequencing for the detection of BRAF V600E mutations in clinical tumour specimens.
authors.affiliation: Oncology Research Institute, National University of Singapore, Singapore.
authors.forename: Yi Hui
authors.initials: YH
authors.lastname: Tan
authors.forename: Yanqun
authors.initials: Y
authors.lastname: Liu
authors.forename: Kong Weng
authors.initials: KW
authors.lastname: Eu
authors.forename: Pei Woon
authors.initials: PW
authors.lastname: Ang
authors.forename: Wei Qi
authors.initials: WQ
authors.lastname: Li
authors.forename: Manuel
authors.initials: M
authors.lastname: Salto-Tellez
authors.forename: Barry
authors.initials: B
authors.lastname: Iacopetta
authors.forename: Richie
authors.initials: R
authors.lastname: Soong
mesh_terms: D045744:Cell Line, Tumor; D003123:Colorectal Neoplasms, Hereditary Nonpolyposis / Q000175:diagnosis / Q000235:genetics; D004252:DNA Mutational Analysis / Q000191:economics / Q000379:methods*; D006801:Humans; D009154:Mutation; D016133:Polymerase Chain Reaction; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf