What is BRAF?

It is a gene that encodes a serine/threonine-protein kinase involved in the MAPK/ERK signaling pathway, which regulates cell growth, differentiation, and survival. When functioning normally, it contributes to controlled cellular signaling. However, certain mutations—especially in codon 600—can activate the gene constitutively, driving uncontrolled cell proliferation and tumorigenesis.

The most frequently observed mutation is V600E (valine to glutamic acid), which accounts for roughly 79% of BRAF-mutated melanomas. Other clinically relevant mutations include V600K, V600R, and V600M.

Role of BRAF Mutations in Cancer

Mutations in BRAF, particularly at codon 600, are key drivers in various cancers. These alterations lead to constant activation of downstream MAPK signaling, promoting unregulated tumor cell division and resistance to apoptosis. BRAF mutations are implicated in:

• Metastatic melanoma (~50% prevalence)
• Papillary thyroid carcinoma
• Colorectal cancer
• Non-small cell lung carcinoma (NSCLC)
• Hairy cell leukemia
• Adenocarcinoma of the lung

Because BRAF mutations are typically mutually exclusive with KRAS, NRAS, and EGFR mutations, their detection is essential for accurate tumor classification and targeted therapy selection.

About the TRUPCR® BRAF Mutation Kit

The TRUPCR® BRAF Mutation Kit is a CE-IVD assay designed for qualitative detection of mutations in codon 600 of the BRAF gene. Using ARMS PCR (Amplification Refractory Mutation System), the assay enables allele-specific amplification, targeting mutated sequences with high precision even in mixed populations of wild-type and mutant DNA.

Key highlights of the kit include:

• Detection of 7 mutations (including V600E, V600K, V600R, and V600M)
• Internal reference gene to ensure sample and extraction quality
• Use of FAM- and HEX-labeled fluorescent probes
• Compatibility with a broad range of real-time PCR platforms

The kit works on DNA extracted from FFPE, fresh, or frozen tumor tissue, making it highly versatile for clinical settings.

Principle and Workflow

1. DNA Extraction: Isolate genomic DNA from FFPE, fresh, or frozen tumor samples.
2. ARMS PCR Amplification: Specific primers selectively amplify mutated BRAF alleles using allele-specific technology.
3. Real-Time Detection: Mutations are identified via fluorescent signal from FAM and HEX probes.
4. Quality Control: An internal reference gene ensures extraction success and helps avoid false negatives.
5. Result Interpretation: Mutations are reported qualitatively, with potential support from software-based curve analysis (if implemented in lab protocol).

The assay is optimized for routine diagnostic use and offers a simple, fast workflow with results available within a few hours.

Clinical Application

Detection of BRAF mutations is central to precision oncology. Identifying codon 600 mutations informs prognosis and eligibility for targeted therapies such as:

• BRAF inhibitors (e.g., vemurafenib, dabrafenib)
• MEK inhibitors (e.g., trametinib)
• Combination therapies in melanoma and NSCLC

This makes the TRUPCR® Mutation Kit ideal for:

• Hospital-based molecular diagnostic labs
• Oncology-focused research labs
• Routine mutation screening in solid tumors

With its CE-IVD status, the kit is validated for clinical use and supports personalized treatment strategies in cancer management.