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

D S Mikhailenko1,2, G D Efremov3, N Yu Safronova3

  • 1N. A. Lopatkin Research Institute of Urology and Intervention Radiology, Affiliated Department of National Medical Research Radiological Center, Ministry of Health of the Russian Federation, Moscow, Russia. dimserg@mail.ru.

Bulletin of Experimental Biology and Medicine
|January 17, 2017
PubMed

Insights

Detecting minor mutations in KRAS, NRAS, and BRAF genes is crucial for cancer diagnosis. Combining real-time PCR and Sanger sequencing offers an optimal algorithm for sensitive and accurate detection of these genetic alterations.

Area of Science:

  • Oncology
  • Molecular Genetics
  • Cancer Genomics

Background:

  • KRAS, NRAS, and BRAF gene mutations are critical in melanoma and colorectal cancer progression.
  • Accurate detection of both common and rare mutations is essential for effective diagnosis and treatment strategies.

Purpose of the Study:

  • To develop an optimal molecular genetic analysis algorithm for detecting minor mutations in KRAS, NRAS, and BRAF genes.
  • To evaluate the sensitivity and accuracy of combined real-time PCR and sequencing methods.

Main Methods:

  • Molecular genetic analysis of KRAS, NRAS, and BRAF genes.
  • Analysis of 35 melanoma and 33 colorectal cancer specimens.
  • Utilized real-time PCR and Sanger sequencing techniques.

Main Results:

  • Frequent KRAS mutations (G12D/V/A/C/S) were identified.
  • The BRAF V600E mutation was most common in melanoma, with a significant percentage (24%) of rare mutations.
  • Sanger sequencing was essential for identifying rare BRAF and NRAS mutations.

Conclusions:

  • A combined approach of real-time PCR and sequencing enhances the sensitivity of mutation detection.
  • This integrated method ensures concordance with international recommendations for genetic analysis in cancer.
Keywords:
oncogenesequencingsomatic mutation

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