DiaCarta's QClamp® BRAF Codon Specific Mutation Detection kit is a polymerase chain reaction (PCR)-based assay that uses XNA probes to suppress amplification of WT-DNA, selectively amplifying mutant DNAs

The testing procedure involves four (4) simple steps:1. Isolation of DNA from tumor biopsies, paraffin-embedded sections (FFPE), fresh frozen tumors, or tumor cell lines2. Amplification of mutant all DNAs at specified codons within the KRAS gene 3. Detection of amplification signal using a real-time PCR instrument capable of SYBR green detection 4. Documentation and interpretation of results. This test can be completed in approximately 2-3 hours from DNA to test result. Limit of Detection <0.1% RUO, <1% CE-IVD. Includes WT and Positive Controls. 2 reactions per sample. Minimal sample input (5-10ng per reaction)

The BRAF gene encodes a serine/threonine protein kinase, which plays a role in regulating the MAP Kinase/ERK signaling pathways, affecting cell growth and proliferation.

BRAF mutations occur in ~50% of melanoma tumors, ~40% of papillary thyroid tumors, ~30% of ovarian tumors, ~10% of colorectal tumors and ~10% of prostate tumors. Drugs that block oncogenic BRAF signaling have had impressive results in clinical trials and the search for new BRAF-targeted drugs is an active area of research and development.

The most common mutations in BRAF occur in codon 600, where an amino acid substitution in the activation segment of the kinase domain creates a constitutively active form of the protein. BRAF V600E accounts for 70-90% of all BRAF mutations identified. Less commonly identified mutations include V600K 10-15%, V600D <5%, V600R <5%, V600M < 1%, and V600E2 <1%. BRAF mutations are generally found in tumors that are wild-type for K-Ras, N-Ras, and EGFR.

The assay identifies the presence of all mutations in and near BRAF Codon 600. All mutations at a particular codon are detected, but the exact nature of the mutation is not specified.