Detection of Rare Mutations by Routine Analysis of KRAS, NRAS, and BRAF Oncogenes

• Published in: Bulletin of experimental biology and medicine Vol. 162; no. 3; pp. 375 - 378
• Main Authors: Mikhailenko, D. S., Efremov, G. D., Safronova, N. Yu, Strelnikov, V. V., Alekseev, B. Ya
• Format: Journal Article
• Language: English
• Published: New York Springer US 2017; Springer; Springer Nature B.V
• Subjects: Algorithms; Base Sequence; Biomedical and Life Sciences; Biomedicine; Cell Biology; Colorectal cancer; Colorectal carcinoma; Colorectal Neoplasms - diagnosis; Colorectal Neoplasms - genetics; Colorectal Neoplasms - pathology; DNA Mutational Analysis; DNA sequencing; Exons; Gene mutation; Genes; Genetic analysis; Genetic aspects; GTP Phosphohydrolases - genetics; Humans; Internal Medicine; Introns; K-Ras protein; Laboratory Medicine; Melanoma; Melanoma - diagnosis; Melanoma - genetics; Melanoma - pathology; Membrane Proteins - genetics; Mutation; Neoplasm Proteins - genetics; Oncogenes; Paraffin Embedding; Pathology; Polymerase chain reaction; Proto-Oncogene Proteins B-raf - genetics; Proto-Oncogene Proteins p21(ras) - genetics; Real-Time Polymerase Chain Reaction; Skin Neoplasms - diagnosis; Skin Neoplasms - genetics; Skin Neoplasms - pathology; Tissue Fixation; oncogene; somatic mutation; sequencing
• ISSN: 0007-4888; 1573-8221
• DOI: 10.1007/s10517-017-3619-z

Molecular genetic analysis of
KRAS
,
NRAS
, and
BRAF
genes was carried out in order to develop an optimal algorithm for detection of minor mutations. We analyzed 35 melanoma and 33 colorectal cancer specimens. Frequent G12D/V/A/C/S mutations were detected in
KRAS
. The most frequent
BRAF
mutation in melanoma was V600E, the percentage of rare mutations is significant for DNA diagnosis (24%). Identification of rare
BRAF
mutations 1790C→G (L597R), 1798_1799delinsAA (V600K), 1798_1799delinsAG (V600R), and 1799_1800delinsAA (V600E) and
NRAS
mutation 38G→T (G13V) was possible only by Sanger sequencing. The combination of real-time PCR and sequencing can improve analysis sensitivity and ensure concordance of the tested loci with the international recommendations.