KRAS/BRAF Mutation Analysis Kit

KRAS/BRAF Mutation Analysis Kit

Available Products

Product Name/Description No. of Reactions* Product Code
KRAS Mutation Analysis Kit for Real-Time PCR (exons 2, 3 and 4) 50 KRAS-RT50
KRAS/BRAF Mutation Analysis Panel Kit for Real-Time PCR (exons 2, 3 and 4 of KRAS and V600E of BRAF) 50 KRBR-RT50
RAS c.59/117 Mutation Detection Kit † 50 RASX-RT50

*Includes all controls

† For detection of KRAS c.59, NRAS c.59/117 somatic mutations. Available with purchase of NRAS-RT50, KRAS-RT50, or KRBR-RT50.

KRAS/BRAF Mutations and Cancer

The KRAS gene encodes a small GTPase that plays a key role in transducing signals from the epidermal growth factor receptor (EGFR) to downstream effectors. KRAS mutations have been commonly found in several types of human malignancies, such as metastatic colorectal cancer (mCRC), lung adenocarcinoma and thyroid cancer. The most common mutations are found in codons 12, 13 and 61. Several studies have demonstrated that tumors carrying any of these mutant forms of the KRAS gene are less likely to respond to anti-EGFR antibody therapy. The American Society of Clinical Oncology (ASCO) recently released its first Provisional Clinical Opinion (PCO) suggesting that all patients to be administered anti-EGFR monoclonal antibody therapy (e.g. cetuximab, panitumumab and erlotnib) should be screened for KRAS mutations. Recent studies have also shown that not all mCRC patients with wild-type KRAS tumors respond to anti-EGFR therapy. This suggests that additional genes and/or pathways may be involved in the mechanism of resistance to these drugs. Mutations in BRAF, another downstream effector of the EGF-activated pathway, have been identified in up to 8% of mCRC tumors. Studies with mCRC patients have shown resistance to anti-EGFR therapy in patients with tumors expressing mutated BRAF. Those same individuals also had decreased progression-free (PFS) and overall (OS) survival when treated with EGFR antagonists. These findings strongly suggest that screening for both KRAS and BRAF mutations is necessary to more accurately identify tumor cells that will not respond to anti-EGFR drugs.

The KRBR-RT50 kit detects the following mutations in KRAS:

Exon Mutation Nucleotide Change Amino Acid Change COSMIC ID
2 G12D c.35G>A Glycine (G) to Aspartic acid (D) 521
G12C c.34G>T Glycine (G) to Cysteine (C) 516
G12S c.34G>A Glycine (G) to Serine (S) 517
G12R c.34G>C Glycine (G) to Arginine (R) 518
G12A c.35G>C Glycine (G) to Alanine (A) 522
G12V c.35G>T Glycine (G) to Valine (V) 520
G13D c.38G>A Glycine (G) to Aspartic acid (D) 532
3 Q61H c.183A>C & c.183A>T Glutamine (Q) to Histidine (H) 554 & 555
Q61L c.182A>T Glutamine (Q) to Leucine (L) 553
Q61R c.182A>G Glutamine (Q) to Arginine (R) 552
4 K117N c.351A>C & c.351A>T Lysine (K) to Asparagine (N) 19940 & 28519
K117R c.350A>G Lysine (K) to Arginine (R) 1178068
K117E c.349A>G Lysine (K) to Glutamic acid (E) 1360831
A146T c.436G>A Alanine (A) to Threonine (T) 19404
A146P c.436G>C Alanine (A) to Proline (P) 19905
A146V c.437C>T Alanine (A) to Valine (V) 19900


Exon Mutation Nucleotide Change Amino Acid Change COSMIC ID
15 V600E c.1799T>A Valine (V) to Glutamic acid (E) 476

For expanded coverage, the RAS c.59/117 Mutation Detection Kit detects KRAS c.59, NRAS c.59/117 somatic mutations.

Testing Procedure and Analysis

EntroGen’s KRAS/BRAF mutation panel is a polymerase chain reaction (PCR)-based assay and uses allele-specific probes to identify the presence of mutations in KRAS exons 2, 3 and 4 as well as BRAF V600E. The testing procedure involves four (4) simple steps:

  • Isolation of DNA from tumor biopsies, paraffin-embedded sections (FFPE), fresh frozen tumors, or tumor cell lines.
  • Amplification of regions of the KRAS and BRAF genes using allele-specific probes.
  • Detection of amplification signal using real-time PCR instrument or gel documentation system.
  • Documentation and interpretation of results.

This test can be completed in approximately 2 hours from DNA to test result.

Equipment and Materials

This assay requires a real-time thermal cycler capable of detecting FAM and VIC fluorescence signals. This test includes reagents required for the PCR amplification and signal detection, as well as validated reaction controls. Columns and reagents for DNA isolation are not included.

Intended Use

USA: EntroGen’s KRAS/BRAF mutation panel reagents are provided for research use only (RUO). Not for use in diagnostic procedures.

Europe: EntroGen’s KRAS/BRAF mutation panel reagents are available for research (RUO) and diagnostic(CE-IVD) purposes.

Bibliography of studies using this product

Poster presented at the 20th National Pathology Congess, Turkey, September 2010. Comparison of K-RAS Mutation Analysis Tests. Veysel Sabri Hançer, Gökhan Demir, İlknur Türkmen, Murat Büyükdoğan, Nuray Başsüllü, Tuncay Altuğ, Reyhan Diz-Küçükkaya, Gülen Bülbül-Doğusoy. Hancer VS, Buyukdogan M, Türkmen I, Bassullu N, Altug T, Diz-Kucukkaya R, Bulbul-Dogusoy G, Demir G. Comparison of KRAS Mutation Tests in Colorectal Cancer Patients. Genet Test Mol Biomarkers. 2011 Jun 23. Pubmed ID 21699410Lewandowska MA, Jóźwicki W, Zurawski B. KRAS and BRAF Mutation Analysis in Colorectal Adenocarcinoma Specimens with a Low Percentage of Tumor Cells. Mol Diagn Ther. 2013 Apr 20. Pubmed ID 23606169 Nicoś M1, Krawczyk P, Jarosz B, Sawicki M, Szumiłło J, Trojanowski T, Milanowski J. Analysis of KRAS and BRAF genes mutation in the central nervous system metastases of non-small cell lung cancer. Clin Exp Med. 2015 Apr 23. Pubmed ID 25902737


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