BRAF Mutations

single page view

General Information

Lab Name
BRAF Mutations
Lab Code
BRAF
Epic Ordering
BRAF Mutations
Description

This test detects the V600E and V600K mutations in the BRAF gene, somatic mutations that occur in about half of melanomas and in 10-15% of colon cancers.

  • BRAF V600E mutations are associated with resistance to antibody therapy against the EGF receptor, and testing may be ordered as a reflex or add-on if the KRAS test is negative.
  • BRAF V600E and V600K mutation status is evaluated to guide treatment with drugs that target BRAF, especially in patients with melanoma.
  • BRAF V600E mutation status is used to rule out Lynch syndrome (HNPCC) in patients with microsatellite unstable cancer.

Please note that BRAF Mutations is intended for solid tumors, for testing related to hematologic malignancies, please order - Heme Single Gene by NGS [HCAPSG]. Consultation with a Director can be requested to determine the appropriate testing. Please contact the laboratory at 206-598-6429 for further questions.

This test may be ordered as a reflex or add-on test.

References
  • Di Nicolantonio F, et al. Wild-type BRAF is required for response to panitumumab or cetuximab in metastatic colorectal cancer. J Clin Oncol 2008, 26:5705-12. 19001320
  • Pritchard CC and Grady WM. Colorectal cancer molecular biology moves into clinical practice. Gut 2011, 60:116-29. 20921207
  • Sharma SG and Gulley ML. BRAF mutation testing in colorectal cancer. Arch Pathol Lab Med 2010, 134:1225-8. 20670148
  • Flaherty KT, et al. Inhibition of mutated, activated BRAF in metastatic melanoma. N Engl J Med 2010, 363:809-19. 20818844
  • Rubinstein JC, et al. Incidence of the V600K mutation among melanoma patients with BRAF mutations, and potential therapeutic response to the specific BRAF inhibitor PLX4032. J Transl Med 2010, 8:67. 20630094
Forms & Requisitions

Genetics Requisition

Synonyms
BRAF gene, DNA, GYNPTH, Gynecological Oncology Pathway, KRAS gene, V600E mutation, V600K mutation, cetuximab, colon cancer, colorectal cancer, melanoma, panitumumab, thor, thorplex, vemurafenib
Components

Interpretation

Method

Next-generation sequencing. Exon 15 of the BRAF gene is captured and sequenced using next-generation sequencing to detect the V600E (GTG to GAG), V600K (GTG to AAG), and other clincally significant BRAF mutations. The test can normally detect a heterozygous mutation if it is present in more than about 5% of the cells in the sample. This test was developed and its performance characteristics determined by the Department of Laboratory Medicine at the University of Washington.

Reference Range
See individual components
Ref. Range Notes

No mutations detected.

References
  • Di Nicolantonio F, et al. Wild-type BRAF is required for response to panitumumab or cetuximab in metastatic colorectal cancer. J Clin Oncol 2008, 26:5705-12. 19001320
  • Pritchard CC and Grady WM. Colorectal cancer molecular biology moves into clinical practice. Gut 2011, 60:116-29. 20921207
  • Sharma SG and Gulley ML. BRAF mutation testing in colorectal cancer. Arch Pathol Lab Med 2010, 134:1225-8. 20670148
  • Flaherty KT, et al. Inhibition of mutated, activated BRAF in metastatic melanoma. N Engl J Med 2010, 363:809-19. 20818844
  • Rubinstein JC, et al. Incidence of the V600K mutation among melanoma patients with BRAF mutations, and potential therapeutic response to the specific BRAF inhibitor PLX4032. J Transl Med 2010, 8:67. 20630094
Guidelines

Ordering & Collection

Specimen Type
Tumor Tissue, Purified DNA, accompanied by a PATHOLOGY REPORT for the tested tissue.
Collection

Requirements for Specimen Selection

  • To ensure clinically relevant results, the most recent and/or metastatic sample is preferred to older specimens, provided sufficient tumor is present (see point 2).
  • To ensure detection of all types of mutations there should be at least 10% tumor cells in the tissue area processed for DNA for mutation detection and 20% tumor cells for microsatellite instability evaluation. If there is more than one tissue block, please provide the block that has the greatest percentage of neoplastic nuclei.
  • Tissue samples and pathology reports will be reviewed by directors upon receipt for acceptability prior to testing. Director consultation for tissue selection is available if needed (contact Genetics lab).

Specimen Types

Tissue samples

Send one of the following:

  1. Slides: 1 slide at 4-micron thickness stained with hematoxylin-and-eosin (H&E) AND 10 unstained, non-baked slides at 10-micron thickness (a minimum of 5 unstained slides is acceptable). Unstained slides can be on charged or uncharged slides.
  2. Tissue Blocks: Provide complete formalin-fixed tissue block containing tumor tissue. Tissue block will be returned at completion of testing.
  3. Fresh/frozen tissue: 5 microgram tissue in cell culture medium or frozen tissue stored at -20C. Tumor percentage will not be determined prior to sequencing studies.

NOTE: In order to ensure that enough DNA is obtained, the minimum acceptable tissue area is 10 square millimeters when ten 10-micron slides are supplied (1 cubic millimeter of tissue).

Purified DNA

5 micrograms ANDa reference hematoxylin-and-eosin (H&E) stained slide and pathology report required.

Bone Marrow

1 to 2 mL Bone Marrow in LAVENDER TOP (EDTA) tube

Blood

6 mL blood in LAVENDER TOP (EDTA) tube.

Alternative specimens may be acceptable with approval (contact: 206-598-1149).

For ADD-ON after prior testing, contact Genetics lab.

Unacceptable samples

We cannot accept decalcified samples or tissue samples treated with fixatives other than formalin.

Quantity:

Requested:

  • Tissue: 10 unstained slides (10-micron thickness) plus one H&E-stained slide.
  • Extracted DNA: 5 microgram Bone Marrow: 2 mL
  • Blood: 6 mL

Minimum:

  • Tissue: 5 unstained slides (10-micron thickness) plus one H&E-stained slide.
  • Extracted DNA: 100-250 nanograms Bone Marrow: 1 mL
  • Blood: 3 mL
Forms & Requisitions

Genetics Requisition

Handling Instructions

Attach a copy of the pathology report for the tumor sample being submitted.

Hold slides or tissue blocks at room temperature.

Outside Laboratories: Ship at room temperature.

Stability: unstained slides or tissue blocks stable at room temperature for at least 2 years.

Quantity
requested: Amounts as noted above
minimum: Amounts as noted above

Processing

Processing

Hold slides or tissue blocks at room temperature.

Outside Laboratory: Ship at room temperature.

Stability: unstained slides or tissue blocks stable at room temperature for at least 2 years

Performance

LIS Dept Code
Genetics (GEN)
Performing Location(s)
UW-MT Genetics

Attention: Genetics Lab
Clinical lab, Room NW220
University of Washington Medical Center
1959 NE Pacific Street
Seattle, WA 98195

Tel: 206-598–6429 M–F (7:30 AM–4:00 PM)
Fax: 206-598–0304
Lab email: genelab@uw.edu

Tel (EXOME only): 206-543-0459

Manager

Rebecca Gaulin, rgaulin@uw.edu

Genetic Counselors

Angela Jacobson, MS, LGC agibson@uw.edu
Sarah Paolucci, MA, MS, LGC, spaolucc@uw.edu
Jenna Huey, MS, LGC, jlhuey@uw.edu
Sandra Coe, MS,LGC, scoe20@uw.edu
Dru Leistritz, MS, LGC, dru2@uw.edu (EXOME testing only)

Variant Review Scientist

Ankita Jhuraney, PhD

Faculty

Colin C. Pritchard, MD, PhD
Brian H. Shirts, MD, PhD
Christina Lockwood, PhD, DABCC
Stephen Salipante, MD, PhD
Eric Konnick, MD, MS
Niklas Krumm, MD,PhD
Vera Paulson, MD, PhD
Jillian Buchan, PhD, FACMG

Frequency
Run at least once a week; results in 2 - 3 weeks from specimen receipt
Available STAT?
No

Billing & Coding

CPT codes
81210
Billing Comments

For pricing information, contact Client Support Services 206-520-4600 or 800-713-5198.

LOINC
58483-9