The QClamp® BRAF Mutation Detection Test for plasma and Formalin-Fixed Paraffin-Embedded (FFPE) samples aids in the identification of patients eligible for cancer treatment and in monitoring response to therapy, which can lead to improved outcomes in cancer patients. The QClamp® BRAF Mutation Detection Test is an in vitro diagnostic real-time qualitative PCR assay for the detection of somatic mutations in and near codon 600 on exon 15, in the BRAF serine/threonine protein kinase gene, using purified DNA extracted from FFPE or plasma.
Powered by XNA technology, the QClamp® BRAF Mutation Detection Test has achieved a much higher analytical sensitivity compared to other commercial qPCR kits and other cancer gene mutation detection methods. QClamp® BRAF Mutation Detection Test is able to detect reliably 0.1% to 0.5% mutant DNA out of wild-type DNA for targeted mutations, providing lower detection limit compared to similar assays available in the market due to robust enrichment of mutant sequences while suppressing amplification of wild-type sequences.
QClamp® BRAF Mutation Detection Test is CE/IVD-certified.
Advantages of QClamp® BRAF Mutation Detection Test
Reliably detects 0.1% to 0.5% VAF mutant DNA out of wild-type DNA for targeted mutations
Suitable for plasma and FFPE samples
LOW INPUT DNA
Minimum 5ng input DNA per reaction. Less than 2 tubes of blood (10mL each) needed for cfDNA
Covering all relevant somatic mutations of BRAF oncogene
Less than 4 hours of assay run time
Validated on the most common qPCR machines with minimized variability
BRAF Mutation and Cancer
The B-type Raf Kinase (BRAF) protein is a serine/threonine kinase that has important roles in regulating the MAP kinase/ERK signaling pathways, affecting cellular proliferation, differentiation, and programmed cell death. A BRAF mutation is commonly found in many human cancers including malignant melanoma, colorectal cancer, lung cancer, papillary thyroid carcinoma, non-small cell lung carcinoma, and adenocarcinoma of the lung.
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. The V600E and V600K mutations are found in high frequencies in human cancer at 70-90% and 10-15%, respectively. BRAF mutations are generally found in tumors that are wild-type for KRAS, NRAS, and EGFR. Therefore, BRAF c600 mutation may serve as a biomarker for diagnosis, prognosis, and treatment options for cancer patients.
BRAF V600E and Targeted Cancer Therapy
Drugs, such as RAF inhibitor vemurafenib inhibit the ERK pathway and cell proliferation and lead to 80% response rate in metastatic melanoma patients carrying the BRAF V600E mutation. However, these drugs do not have therapeutic effects for colorectal cancer patients carrying the same mutation. This is due to overexpression of EGFR in BRAF V600E colorectal cancer cells, but not in melanoma cells because they overcome the inhibition of the BRAF mutant kinase activity. In these colorectal cancer patients, use of tyrosine kinase inhibitors such as gefitinib, erlotinib, and cetuximab together with vemurafenib has a synergistic effect.
Supporting Data for QClamp® BRAF Mutation Detection Test
QClamp® BRAF Mutation Detection Test analytical sensitivity in plasma: detects as low as 0.1% VAF mutant DNA in a 5ng input
cfDNA reference material from SeraCare with known BRAF V600E VAF was diluted to 1%, 0.5% and 0.1% VAF with 2.5ng/μl of wild-type cfDNA. The BRAF V600E assay was able to detect as low as 1-2 copies of BRAF V600E mutant (0.1% VAF) in the background of wild-type cfDNA.
QClamp® BRAF Mutation Detection Test analytical sensitivity in FFPE: detects as low as 0.1% VAF mutant DNA in a 5ng input.
Genomic DNA (gDNA) reference material from Horizon Discovery with known BRAF V600E VAF was diluted to 1%, 0.5% and 0.1% VAF with 5 ng/μl wild-type cfDNA. The BRAF V600E assay was able to detect as low as 3-4 BRAF V600E mutant copies (0.1% VAF) in the background of wild-type gDNA.
Streamlined Workflow for QClamp® Gene Mutation Detection Tests
Step 1: DNA Isolation & Quantification
Extract DNA from FFPE or plasma using a commercial DNA extraction kit and measure the concentration using fluorometric analysis
Step 2: set up qpcr
Mix the assay reagents, load into PCR plate, add controls and extracted DNA ~ 30-60 minutes
Step 3: Amplification parameters
Enter amplification parameters on
qPCR instrument, load PCR plate
and start the run ~ 2.5 hours
Step 4: Data analysis
Determine the presence or absence
of mutations according to the Cq
value cutoffs ~ 15 minutes
CE/IVD catalog #: DC-10-0197
Research-use-only (RUO) catalog #: DC-10-0197R
In vitro diagnostic Use (CE/IVD) or for research use
Plasma and FFPE
Roche LightCycler® 480, Bio-Rad CFX384 and ABI QuantStudio 5
Less than 4 hours
Stable for 12 months at -25 ℃ to -15 ℃
Most frequent BRAF mutations detected by QClamp® BRAF Mutation Detection Test
|Exon||Amino Acid Change||Cosmic No.|
|15||p.V600E||COSM476, COSM475, COSM1131|
For products that are in stock, DiaCarta will arrange shipment in 1-3 days. For products that are on backorder, DiaCarta will arrange shipment in 3-5 weeks.
Intended Use: QClamp® BRAF Mutation Detection Test is CE/IVD-certified. Outside
Shipping Condition: QClamp® BRAF Mutation Detection Test will be shipped with dry ice. For domestic shipment, DiaCarta provides overnight delivery through FedEx Domestic Overnight Shipping Service. For international shipment, DiaCarta provides 3-7 days in transit through FedEx International Priority Shipping Service. Please contact DiaCarta if you prefer to use your own shipping carrier.