Detection of c-kit-activating mutations in gastrointestinal stromal tumors by high-resolution amplicon melting analysis

Am J Clin Pathol. 2004 Aug;122(2):206-16. doi: 10.1309/4E6U-YBY6-2N2F-CA6N.


High-resolution amplicon melting analysis was used to scan for c-kit-activating mutations in exons 9, 11, 13, and 17 in 29 neoplasms diagnosed as gastrointestinal stromal tumors (GISTs). Immunohistochemically, 7 of 29 did not show strong CD 17 positivity and might represent true smooth muscle tumors or c-kit-negative GISTs. No c-kit-activating mutations were detected in the 7 CD117- cases by high-resolution amplicon melting analysis or direct DNA sequencing. Alterations in the remaining 22 CD117+ cases included 13 (59%) in exon 11, 2 (9%) in exon 9, 1 (5%) in exon 13, and none in exon 17. The genetic alterations consisted of point mutations and in-frame insertions, duplications, and deletions. In exon 11, 7 (54%) of 13 alterations have not been described previously. In 2 cases, the identical exon 11 mutation was observed in the primary tumor and a metastatic/recurrent lesion. In all cases, direct DNA sequencing confirmed that polymerase chain reaction products with an abnormal melting curve contained a mutation and products with a normal melting curve, a normal DNA sequence. High-resolution melting analysis can be used to scan DNA for potential c-kit-activating mutations and can aid in the diagnosis of GISTs.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • DNA Mutational Analysis
  • DNA Primers
  • DNA Topoisomerases, Type II / biosynthesis
  • Gastrointestinal Neoplasms / genetics*
  • Gastrointestinal Neoplasms / pathology
  • Humans
  • Immunohistochemistry
  • Molecular Sequence Data
  • Mutation
  • Nucleic Acid Amplification Techniques* / methods
  • Polymerase Chain Reaction
  • Proto-Oncogene Proteins c-kit / genetics*
  • Stromal Cells / pathology*
  • Transition Temperature


  • DNA Primers
  • Proto-Oncogene Proteins c-kit
  • DNA Topoisomerases, Type II