Lab Test

Test Definition Number : 71
Test Definition Name : BRAF sequence analysis
Lab Name : OHSU Knight Diagnostics Lab
Lab Test Number of THIS Version : 1175
Lab Test Name : BRAF Mutation Analysis
Status : Active
Expired : No
Updated : 2018-01-27 21:52 EST

General Attributes

Chromosome Band : 7q34
HUGO Gene Name : [BRAF] B-Raf proto-oncogene, serine/threonine kinase
ICD-Oncology Code
(includes WHO classification diagnoses)
:

Infectious Disease Attributes

Agent Category :
Non-virus Genus and Species :
Virus Family, Genus, and Species :

Lab-specific Attributes

Method : Polymerase Chain Reaction (PCR) - Real Time
Sequencing - Sanger - Bi-directional
Specimen Source
(for clinical patient care these sources must be validated)
: Blood - Whole Blood
Bone Marrow Aspirate
Solid Tissue Paraffin-Embedded Formalin-Fixed
ICD-10 Code :
Use :
Genetics Category : Acquired (somatic)
SNOMED CT :
LOINC :
Comments : Background: Melanoma - Mutations in the BRAF gene are the most common oncogenic alterations in malignant melanoma, present in approximately 45% of tumors arising from cutaneous sites and also occurring in melanomas at other locations. The V600E mutation is the most common BRAF alteration, and this mutation is the target for BRAF kinase inhibitors such as vemurafenib. Colorectal adenocarcinoma - BRAF gene mutations are present in approximately 12% of cases and correlate with resistance to treatment with cetuximab. In addition, BRAF mutations are a significant negative prognostic factor in patients with stage II and III colorectal carcinoma. Methodology: 1. Microscopic examination of the specimen and macrodissection of tumor-rich areas. 2. DNA extraction and purification. 3. PCR amplification of BRAF exon 15. 4. Screening for mutations by one of two methods, depending on tumor cellularity. a. Real-time PCR with high resolution melting curve analysis (HRM). DNA sequencing is used to confirm any potential mutations identified by this approach. b.Bidirectional Sanger sequencing using an LNA-modified oligonucleotide to suppress wild-type allelic amplification (LNA-sequencing). 5. Estimated sensitivity: 6. 20% mutant allele for HRM. 7. 1% mutant allele for LNA-sequencing (used for samples with low tumor cellularity) 8. Estimated specificity: 99.5% of BRAF mutations reported in melanoma and colorectal carcinoma.
Specimen Transport Requirements : 5-10mL Blood or Bone Marrow (ACD or EDTA) Paraffin block or 10 unstained sections (4-5 micron) Due to DNA damage, decalcified specimens are not recommended. Package and ship specimens to remain cold, but not frozen. Ship via overnight express. Contact Client Services at (855) 535-1522 for shipping kits and instructions.
Samples Available for Proficiency Test Sample Exchange : Yes

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