BRAF Mutation Detection Test | DiaCarta, Inc.

To aid in the identification of patients eligible for treatment and to monitor response to therapy, DiaCarta offers QClamp® BRAF Mutation Detection Test in plasma and Formalin-Fixed Paraffin-Embedded (FFPE) samples 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.

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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 with competition due to a strict 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

ULTRA-SENSITIVE

Detect reliably 0.1% to 0.5% VAF mutant DNA out of wild-type DNA for targeted mutations

SAMPLE READY

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

COMPREHENSIVE COVERAGE

Covering all relevant somatic mutations of BRAF oncogene

FAST RESULTS

Less than 4 hours of assay run time

GREAT VERSATILITY

Validated on the most common qPCR machines with minimized variability

BRAF Mutation and Cancer

BRAF Introduction

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 lung.
 
 
 

BRAF Mutations

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, that overcomes 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 synergistic effect. 

Streamlined Workflow for QClamp® BRAF Mutation Detection Tests

Step 1: DNA Isolation & Quantification

Extract DNA from FFPE or plasma using a commercial DNA extraction kit followed by measuring 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

Resources

Catalog Number 

CE/IVD catalog #: DC-10-0197
Research-use-only (RUO) catalog #: DC-10-0197R

Pack Size

30 samples

Detected Codons

Codon 600

Intended Use

In vitro diagnostic Use (CE/IVD) or for research use

Sample Type

Plasma and FFPE

Input DNA

5-10ng/Reaction

Validated Instruments

Roche LightCycler® 480, Bio-Rad CFX384 and ABI QuantStudio 5

Detection Channel

FAM; HEX

Detection Chemistry

TaqMan

Turnaround Time

Less than 4 hours

Stability

Stable for 12 months at -25 ℃ to -15 ℃

Most frequent BRAF mutations detected by QClamp® BRAF Mutation Detection Test

ExonAmino Acid ChangeCosmic No.
15p.V600ECOSM476, COSM475, COSM1131
p.V600KCOSM473, COSM26487
p.V600RCOSM474, COSM21617
p.V600DCOSM477, COSM36128

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 is able to detect as low as 1-2 copies of BRAF V600E mutant (0.1% VAF) in the background of wild-type cfDNA.

 

Analytical sensitivity in gDNA (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 is able to detect as low as 3-4 BRAF V600E mutant copies (0.1% VAF) in the background of wild-type gDNA.

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Ordering Information

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. For the outside USA, this product is available for diagnostic use (CE/IVD) and research use (RUO). For the USA, this product is provided for research use only (RUO) and not for diagnostic use. DiaCarta ships CE/IVD version to the outside USA and ships RUO version to the USA. Please call 1-800-246-8878 or email order@diacarta.com if you have questions or specific needs.

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.

Need Help?

Call us: +1 (800) 878-6662

Email us: information@diacarta.com

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