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Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines
Published on: October 8, 2015
Molecular platforms utilized to detect BRAF V600E mutation in melanoma
Jonathan L Curry1, Carlos A Torres-Cabala, Michael T Tetzlaff
1Department of Pathology, Section of Dermatopathology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA. jlcurry@mdanderson.org
Seminars in Cutaneous Medicine and Surgery
|November 24, 2012
Summary
Metastatic melanoma (MM) treatment is improved by identifying the BRAF V600E mutation. The cobas BRAF V600 Mutation Test offers a highly sensitive method for detecting this key mutation in tumor samples.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Metastatic melanoma (MM) is a deadly skin cancer with unpredictable clinical outcomes.
- Identifying key biomarkers is crucial for understanding melanoma's aggressive behavior and developing targeted therapies.
- The BRAF V600E mutation is a common driver of melanoma cell proliferation.
Purpose of the Study:
- To highlight the importance of molecular signatures in guiding melanoma patient management.
- To evaluate the performance of diagnostic platforms for BRAF mutation analysis.
- To assess the utility of the cobas BRAF V600 Mutation Test for clinical application.
Main Methods:
- Review of existing molecular platforms for melanoma mutation analysis, including Sanger sequencing, pyrosequencing, and next-generation sequencing.
- Focus on the cobas BRAF V600 Mutation Test for detecting BRAF V600E mutations in formalin-fixed paraffin-embedded tumor samples.
- Comparison of the cobas test's analytic performance with Sanger sequencing and 454-pyrosequencing.
Main Results:
- The cobas BRAF V600 Mutation Test demonstrated >99% sensitivity in detecting BRAF V600E mutations compared to Sanger sequencing.
- The test has a low limit of detection (4%-5% mutant alleles) in tissue samples.
- High sensitivity and reproducibility were confirmed for BRAF V600E mutation analysis.
Conclusions:
- Knowledge of the molecular signature, particularly BRAF V600E status, is vital for directing melanoma treatment.
- The cobas BRAF V600 Mutation Test is a sensitive and reproducible assay for clinical use.
- Targeted therapies like BRAF inhibitors have revolutionized MM treatment, emphasizing the need for accurate mutation detection.
