|BRAF Mutation Analysis, Tissue [BRAF]|
| WSLH Department:|| Cytogenetics|
|WSLH Test Code:||840C41|
|Includes:||Mutations in the BRAF gene have been reported in many human cancers, including malignant melanoma (27-70%), colorectal cancer (5-22%), ovarian cancer (30%) and papillary thyroid cancer (30-70%). This assay detects the predominant BRAF mutation, a single transversion in exon 15 (T1799A) resulting in an amino-acid substitution of glutamate for valine at residue 600 (Val600Glu). Consideration of BRAF status should be given when determining appropriate therapy.
Specific BRAF kinase inhibitors, such as Zelboraf, have recently gained FDA approval for BRAF V600E positive metastatic melanoma. Furthermore, the BRAF V600E mutation is a known negative predictor of Cetuximab therapy.|
|Methodology:||**(CPT Code: 81210)** Real-Time PCR followed by Melting Point Analysis|
|Availability:||Once a week.|
|Turn-around Time:||10 Days|
|Recommended Uses:||Formalin-fixed, paraffin embedded tissue or fine needle aspirates (FNA)|
|Specimen Requirements:||Formalin-fixed, paraffin embedded tissue or fine needle aspirates (FNA)|
|Collection Kit/Container:|| |
|Patient Preparation:|| |
|Collection Instructions:||Please fill out Clinical Laboratories Referral Screening Flow Sheet if test is being added to specimen that was collected >30 days before add on order or call UWHC Test Referral office at (608) 262-6388.|
|Specimen Handling and Transport:||Three slides each containing 5 microns (uM) of FFPE tissue should be sent. Second slide should be H&E stained with the tumor circled. Please indicate percent tumor on Intra-Lab Send-Out Form.
Fine Needle Aspirates (FNA) have also been validated for this method.
If an add-on order is needed, please contact UWHC Surgical Pathology at (608)263-8443. A Surgical Pathology Tissue Examination Request form will need to be completed and faxed to (608)262-7174.|
|Unacceptable Conditions:||Specimens processed in alternative fixatives.|
|Requisition Form:||Cytogenetics Lab Neoplasia Diagnosis Form #132|
|Required Information:||Surg Path submits appropriate specimen (paraffin block or slides) to Core Lab with an Intra-Laboratory Send-Out Order Form. Client Service staff will place sample in Molecular Diagnostics ambient bin. Test will be entered in Horizon and PowerPath by Molecular Diagnostics staff.
a) If collect date is less than 30 days + sample is collected during stay-use IP encounter and actual collect date.
b) If collect date is greater than 30 days, create HOV for receive date. Use received date as date of service, enter actual collect date as a test comment. Use SPATH or indicated Clinic as ordering location/source facility.
a) If collect date is less than 30 days + existing encounter for the same date, se this encounter. No HOV needs to be created.
b) If collect date is less than 30 days + no encounter for that date, create HOV using collect date. Use SPATH or indicated Clinic as ordering location/source facility.
c) If collect date is greater than 30 days, create HOV for receive date. Use received date as date of service, enter actual collect date as a test comment. Use SPATH or indicated Clinic as ordering location/source facility.|
|Results include:||Not detected.
A written interpretive report is provided by the laboratory.|
|Limitations:||A "Not Detected" result does not preclude the presence of V600E mutation in BRAF since analytical detection depends on numerous factors such as: the heterogenous nature of the tissue sample and ~5% analytical sensitivity of this assay, sample integrity and the absence of PCR inhibitors. Additionally, other dinucleotide mutations affecting codon 600 of BRAF have been observed and are not directly tested for with this assay. Moreover, please note this assay does not rule out the presence of other mutations in the BRAF gene or other components of the EGFR signaling cascade.|
|Additional Tests Recommended:|| |
|Additional Comments:||A "Detected" result indicates the presence of a V600E BRAF mutation. A "Not Detected" result does not rule out the presence of a BRAF mutation. Inadequate specimen collection, processing and storage may invalidate test results. Test results should not be used as the only criterion to form a clinical conclusion but should be interpreted in the context of all clinical findings, tumor sampling and laboratory data.
The performance characteristics of this test were validated by UWHC Clinical Laboratories. The U.S. Food and Drug Administration (FDA) has not approved or cleared this test; however, FDA approval or clearance is currently not required for clinical use of this test. The results are not intended to be used as the sole means for clinical diagnosis or patient management decisions. The UWHC Clinical Laboratories is authorized under Clinical Laboratory Improvement Amendments (CLIA) to perform high-complexity testing.|
|Additional Tests Performed:|| |