BRAFm+ Metastatic melanoma
What makes it different, makes it vulnerable

BRAF mutation testing: published evaluations

The different methods for assessing BRAF mutations can be compared based on:

  • Performance characteristics
    • Specificity and sensitivity (analytical and clinical)
    • Reproducibility – between-laboratory precision
    • Repeatability – within-laboratory precision
  • Target: what exactly they detect (number of mutations, which mutations)
  • Regulatory status (i.e. US [FDA], European [CE marking], or country-specific [Laboratory Developed Tests (LDT)])
  • Instrumentation required to run the test
  • Throughput (how many samples can be tested at once and within a given time)
  • Percentage of invalid tests (where no result is generated, which can be due to limitation of the method and/or quality of the sample)
  • Cost of testing and time to result

Below are some of the published results of different expert teams, comparing different methods.

Publication Methods compared
Comparison of Testing Methods for the Detection of BRAF V600E Mutations in Malignant Melanoma: Pre-Approval Validation Study of the Companion Diagnostic Test for Vemurafenib.1 Cobas BRAF Mutation Test; ABI BRAF test; and bidirectional direct sequencing
Comparison of Targeted Next-Generation Sequencing (NGS) and Real-Time PCR in the Detection of EGFR, KRAS, and BRAF Mutations on Formalin-Fixed, Paraffin-Embedded Tumor Material of Non-Small Cell Lung Carcinoma—Superiority of NGS.2 NGS, Real-time PCR
Detection of BRAF V600 mutations in metastatic melanoma: comparison of the Cobas 4800 and Sanger sequencing assays.3 Cobas BRAF Mutation Test; Sanger sequencing
Clinical validation of KRAS, BRAF, and EGFR mutation detection using next-generation sequencing.4 NGS
Validation of Next Generation Sequencing Technologies in Comparison to Current Diagnostic Gold Standards for BRAF, EGFR and KRAS Mutational Analysis.5 NGS, Sanger sequencing, real-time PCR
Comparison of high resolution melting analysis, pyrosequencing, next generation sequencing and immunohistochemistry to conventional Sanger sequencing for the detection of p.V600E and non-p.V600E BRAF mutations.6 HRM, NGS, pyrosequencing, IHC, Sanger sequencing
Comparative evaluation of the new FDA approved THxID™-BRAF test with high resolution melting and sanger sequencing.7 THxID BRAF test Real-time PCR, HRM, Sanger sequencing
KRAS and BRAF mutation analysis in routine molecular diagnostics: comparison of three testing methods on formalin-fixed, paraffin-embedded tumor-derived DNA.8 HRM, real-time PCR
Evaluation of BRAF Mutation Testing Methodologies in Formalin-Fixed, Paraffin-Embedded Cutaneous Melanomas.9 Cobas 4800 BRAF V600 Mutation Test, Sanger sequencing, Pyrosequencing, real-time PCR
Detection of BRAF p.V600E Mutations in Melanomas Comparison of Four Methods Argues for Sequential Use of Immunohistochemistry and Pyrosequencing.10 IHC, Pyrosequencing, Sanger Sequencing, real-time PCR
Implementation of a companion diagnostic in the clinical laboratory: The BRAF example in melanoma.11 HRM, real-time PCR, sequencing, NGS, Cobas 4800 BRAF V600, Biomerieux ThxID BRAF
  1. Lopez-Rios F, et al. Comparison of Testing Methods for the Detection of BRAF V600E Mutations in Malignant Melanoma: Pre-Approval Validation Study of the Companion Diagnostic Test for Vemurafenib. PLoS ONE. 2013;8(1): e53733.
  2. Tuononen K, et al. Comparison of Targeted Next-Generation Sequencing (NGS) and Real-Time PCR in the Detection of EGFR, KRAS, and BRAF Mutations on Formalin-Fixed, Paraffin-Embedded Tumor Material of Non-Small Cell Lung Carcinoma—Superiority of NGS, Genes, Chromosomes & Cancer. 2013;52:503–11.
  3. Qu K, Pan Q, Zhang X, et al. Detection of BRAF V600 mutations in metastatic melanoma: comparison of the Cobas 4800 and Sanger sequencing assays. J Mol Diagn. 2013;15:790–5.
  4. Lin MT, et al. Clinical validation of KRAS, BRAF, and EGFR mutation detection using next-generation sequencing. Am J Clin Pathol. 2014;141(6):856–66.
  5. McCourt CM, et al. Validation of Next Generation Sequencing Technologies in Comparison to Current Diagnostic Gold Standards for BRAF, EGFR and KRAS Mutational Analysis. PLoS ONE. 2013;8(7):e69604.
  6. Ihle MA, et al. Comparison of high resolution melting analysis, pyrosequencing, next generation sequencing and immunohistochemistry to conventional Sanger sequencing for the detection of p.V600E and non-p.V600E BRAF mutations. BMC Cancer 2014;14(13):1–13.
  7. Marchant J, et al. Comparative evaluation of the new FDA approved THxID™-BRAF test with high resolution melting and sanger sequencing. BMC Cancer 2014;14(13):1-13.
  8. Heideman DA, et al. KRAS and BRAF mutation analysis in routine molecular diagnostics: comparison of three testing methods on formalin-fixed, paraffin-embedded tumor-derived DNA. The Journal of molecular diagnostics: JMD. 2012;14(3):247–55.
  9. Lade-Keller J, et al. Evaluation of BRAF Mutation Testing Methodologies in Formalin-Fixed, Paraffin-Embedded Cutaneous Melanomas. J Mol Diagn. 2013;15:70–80.
  10. Colomba E, et al. Detection of BRAF p.V600E Mutations in Melanomas Comparison of Four Methods Argues for Sequential Use of immunohistochemistry and Pyrosequencing. J Mol Diagn. 2013;15:94–100.
  11. Mancini I, et al. Implementation of a companion diagnostic in the clinical laboratory: The BRAF example in melanoma. Clinica Chimica Acta. 2014;439:128–36.