title: Non-p.V600E BRAF Mutations Are Common Using a More Sensitive and Broad Detection Tool.
abstract: OBJECTIVE
  To assess the performance of a next-generation sequencing (NGS) platform for the clinical detection of BRAF mutations.
  
  
  METHODS
  In this retrospective quality assessment of an NGS assay, we analyzed BRAF mutations within parts of exons 11 and 15 in 835 neoplastic tissues submitted to our molecular diagnostics laboratory.
  
  
  RESULTS
  The NGS assays detected a BRAF mutation in 5.9% of lung adenocarcinomas, 13% of colorectal cancers, and 44% of melanomas. Mutant allele frequencies were less than 20% in 28% of 88 BRAF-mutated specimens. Two lymph node specimens with subcapsular or infiltrative metastasis showed 1% to 2% mutant alleles. There were 26 unique BRAF mutations in exons 11 and 15, including three novel mutations. Mutations were located outside codon 600 in 39% of BRAF-mutated tumors. Lung adenocarcinomas showed significantly higher non-p.V600E mutations (86%) than did colorectal cancers (23%) and melanomas (34%). The three most common BRAF mutations in lung cancers accounted for only 41% of the observed BRAF mutations (p.D594G [18%], p.V600E [14%], and p.G469A [9%]).
  
  
  CONCLUSIONS
  The NGS assay demonstrated a high analytic sensitivity and a broad reportable range for clinical detection of BRAF mutations. Elucidating the spectrum of non-p. V600E BRAF mutations in different malignancies is a first step toward understanding their clinical significance.
authors.affiliation: From the Departments of Pathology and.
authors.forename: Jamal
authors.initials: J
authors.lastname: Carter
authors.affiliation: From the Departments of Pathology and Department of Medical Genetics, National Taiwan University Hospital, Taipei; and.
authors.forename: Li-Hui
authors.initials: LH
authors.lastname: Tseng
authors.affiliation: From the Departments of Pathology and.
authors.forename: Gang
authors.initials: G
authors.lastname: Zheng
authors.affiliation: From the Departments of Pathology and Department of Pathology, Massachusetts General Hospital, Boston.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Dudley
authors.affiliation: From the Departments of Pathology and.
authors.forename: Peter
authors.initials: P
authors.lastname: Illei
authors.affiliation: From the Departments of Pathology and Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD;
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Gocke
authors.affiliation: From the Departments of Pathology and Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD;
authors.forename: James R
authors.initials: JR
authors.lastname: Eshleman
authors.affiliation: From the Departments of Pathology and mlin36@jhmi.edu.
authors.forename: Ming-Tseh
authors.initials: MT
authors.lastname: Lin
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D009369:Neoplasms / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D048493:Proto-Oncogene Proteins B-raf; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf
keywords: BRAF; Colorectal cancer; Lung cancer; Melanoma; Next-generation sequencing