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Updated: Aug 24, 2026
Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines
Published on: October 8, 2015
Rapid detection of mutations in the BRAF gene using real-time polymerase chain reaction and melting curve analysis
Tsuneo Ikenoue1, Yohko Hikiba, Fumihiko Kanai
1Division of Gastroenterology, The Institute for Adult Diseases, Asahi Life Foundation, 1-9-14, Nishi-Shinjuku, Shinjuku-ku, Tokyo 160-0023, Japan. t-ikenoue@asahi-life.or.jp
Abstract:
The BRAF gene is mutated in 66% of melanomas and less frequently in various human cancers. More than 80% of these mutations are T to A transversions at nucleotide 1796 (T1796A), leading to a substitution of glutamic acid for valine at amino acid 599 (V599E). We established a new method for rapidly detecting V599E mutations using real-time polymerase chain reaction and melting curve analysis. Furthermore, we examined mutations in gastrointestinal cancer cell lines using this method. We found a mutation in 1 of 12 (8%) colorectal cancer cell lines, but no mutation was detected in 9 gastric cancer cell lines. These results suggest that the BRAF mutation is unlikely to be involved in gastric carcinogenesis.
