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HelpTest Code LAB2851 BRAF Mutations
Clinical System Name
BRAF Mutations
Synonyms
600E mutation, V600E mutation, V600K mutation, BRAF gene
Description
This test detects the V600E and V600K mutations in the BRAF gene. It does not detect fusions.
Sample Requirements
Specimen: Tissue (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.
Note: sections should contain as much tumor as possible.
2. Tissue Blocks: Provide complete formalin-fixed tissue block containing tumor tissue. If there is more than one tissue block, please provide the block that has the greatest amount of 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.
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.
Specimen: Extracted DNA
Container(s): Sterile plastic tube
Preferred Vol: 5 µg AND a reference hematoxylin-and-eosin (H&E) stained slide and pathology report required.
Specimen: Bone Marrow
Container(s): Lavender Top/EDTA
Preferred Vol: 1 to 2 mL Bone Marrow
Specimen: Whole Blood
Container(s): Lavender Top/EDTA
Preferred Vol: 6 mL blood in
Processing Instructions
Reject due to: Decalcified sample
Spin: N
Aliquot: N
Temp: 2 - 8 C
Storage location: Affix labels to the tube(s) and place in the CPA refrigerator, Send Outs rack.
Off-site collection: Send whole blood refrigerated. Optimal results are achieved when samples are received within 72 hours of collection for both bone marrow and peripheral blood
Stability
| Specimen Type | Temperature | Time |
|---|---|---|
| Whole Blood | Room temp | 3 d |
| Refrigerated | 7 d | |
| Frozen | N | |
| Extracted DNA | Room temp | 3-4 d |
| Refrigerated | 1 y | |
| Frozen | Indefinitely |
Availability
| STAT | Performed | TAT |
|---|---|---|
| N | Once a week | 2 - 3 w |
Performing Laboratory
University of Washington
Department of Laboratory Medicine
UW Genetics Lab
1959 NE Pacific St, NW220
Seattle, WA 98195
Phone Number: 206-598–6429
Department
Department: Send Outs/Genetic
Phone Number: (206) 987-2563
Methodology
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
Interpretive report is provided.
Send Out Instructions
| Reference Test Name: | BRAF Mutations |
| Reference Test Number: | BRAF |
| Instructions: | Send out Monday - Friday with the UW courier. |