title: Detecting BRAF Mutations in Formalin-Fixed Melanoma: Experiences with Two State-of-the-Art Techniques.
abstract: BACKGROUND
  Melanoma is characterized by a high frequency of BRAF mutations. It is unknown if the BRAF mutation status has any predictive value for therapeutic approaches such as angiogenesis inhibition.
  
  
  METHODS
  We used 2 methods to analyze the BRAF mutation status in 52 of 62 melanoma patients. Method 1 (mutation-specific real-time PCR) specifically detects the most frequent BRAF mutations, V600E and V600K. Method 2 (denaturing gel gradient electrophoresis and direct sequencing) identifies any mutations affecting exons 11 and 15.
  
  
  RESULTS
  Eighteen BRAF mutations and 15 wild-type mutations were identified with both methods. One tumor had a double mutation (GAA) in codon 600. Results of 3 samples were discrepant. Additional mutations (V600M, K601E) were detected using method 2. Sixteen DNA samples were analyzable with either method 1 or method 2. There was a significant association between BRAF V600E mutation and survival.
  
  
  CONCLUSIONS
  Standardized tissue fixation protocols are needed to optimize BRAF mutation analysis in melanoma. For melanoma treatment decisions, the availability of a fast and reliable BRAF V600E screening method may be sufficient. If other BRAF mutations in exons 11 and 15 are found to be of predictive value, a combination of the 2 methods would be useful.
authors: Department of Dermatology, University Hospital Zürich, Zürich, Switzerland.
authors: Nicola L
authors: NL
authors: Schoenewolf
authors: Reinhard
authors: R
authors: Dummer
authors: Daniela
authors: D
authors: Mihic-Probst
authors: Holger
authors: H
authors: Moch
authors: Mathew
authors: M
authors: Simcock
authors: Adrian
authors: A
authors: Ochsenbein
authors: Silke
authors: S
authors: Gillessen
authors: Peter
authors: P
authors: Schraml
authors: Roger
authors: R
authors: von Moos