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BRAF Extended mutation panel

BRAF Extended Mutation Panel (V600E, V600K & V600R)

  • Clinical Implications
    • Melanoma
      • BRAF mutation is the most common oncogenic driver in melanoma (~ 60%)
      • Many BRAF fusions contribute to different responses to MAPK inhibition
      • BRAF inhibitors
        • Small molecule that binds mutated BRAF and renders protein inactivation
        • FDA-approved inhibitors: Vemurafenib, dabrafenib, trametinib
      • More than 70% of BRAF mutations are V600E and 10-30% are V600K. Potent and selective BRAF inhibitors have demonstrated significant clinical benefits in patients with V600E and V600K BRAF-mutated melanoma.
      • V600R mutations constitute approximately 3-7% of all BRAF mutations.

  • Test Description
  • Real-time PCR for quantitative detection of BRAF V600E, D, R and K mutations.

  • Test approach
  • DNA was isolated and amplified by quantitative polymerase chain reaction (qPCR) using BRAF V600E, D, R and K CE-IVD kit. according to manufacture protocol. Results interpreted according to the manufacture user guide.

  • Reporting name
  • BRAF extended mutation panel

  • Test prerequisites (To ensure timely results)
    • Patient’s demographic data.
    • Clinicopathologic information:
      • Pathology report (final or preliminary) including anatomic location.
      • History of any given therapy for cancer and its date and relation to sample sent for molecular study (i.e. pre & post therapy). Therapy includes chemo and radiotherapy, hormonal or targeted therapy.
      • Any other relevant clinical data or history.
    • Type of sample:
      • Preferred: Formalin-fixed, paraffin-embedded (FFPE) tumor tissue block.
      • Acceptable:
        • Section in Eppendorf: Up to 4 sections, each with a thickness of up to 10 μm and a surface area of up to 250 mm2 + good H&E slide for assessment.
        • Five unstained slides + one good H&E slide.
      • Specimen Minimum Volume: Two 10-micron sections of FFPE.

  • Quality Control measures
  • All samples are subject to stringent quality control measures that include:

    • From your side:
      • Double check you are fulfilling all required data before sending your sample.
      • Check that your pathologist has selected the best block in terms of tumor cellularity, with least presence of necrosis and inflammation.
      • Pretherapy sample is preferred (if underwent any cancer therapy).
    • In our lab:
      • Assessment of tissue for adequacy & tumor cellularity before any molecular analysis.
      • Matching block ID with the report ID and demographic data.
      • Matching the submitted block with the data reported in the pathology report.
    • N.B.
      • If the sample sent in Eppendorf, it is your pathology lab’s responsibility to ensure the sample in Eppendorf is corresponding to the submitted H&E slide (we can’t prepare slide from Eppendorf).
      • This test does not include a pathology consultation.

  • Test Time
  • From 3 days to 5 working days.

  • Retention of the sample
  • Client provided paraffin blocks & unstained slides (if provided) will be returned after testing is complete.

  • Selected References
    1. Turner JA et al: BRAF fusions identified in melanomas have variable treatment responses and phenotypes. Oncogene. ePub, 2018
    2. Ross JS et al: The distribution of BRAF gene fusions in solid tumors and response to targeted therapy. Int J Cancer. 138(4):881-90, 2016
    3. Aramini JM et al: The RAS-binding domain of human BRAF protein serine/threonine kinase exhibits allosteric conformational changes upon binding HRAS. Structure. 23(8):1382-93, 2015