Thesis (Ph.D.) - Cairo University - Faculty of Medicine - Department of Clinical and Chemical Pathology

Background: Thev-raf murine sarcoma viral oncogene homolog B1 (BRAF) is an oncogene and most frequently mutated in RAF family of kinases. Out of BRAF mutations, BRAF p.V600E is the commonest, which can be linked to cancer development especially in melanoma, colon cancer and hairy cell leukemia. BRAF mutation status has emerged as an important predictive marker for eligibility of BRAF inhibitors with promising outcome. High-resolution melting analysis (HRM) can provide sensitive low cost detection of BRAF mutation which sensitivity can be improved when combined with COLD PCR. Aim of the work:Detection of BRAF p.V600E gene mutation in the studied group of hematological malignancy patients using HRM alone, followed by its detection using combined HRM with COLD PCR to enrich minor alleles and comparing both results. Subjects and methods:Thirty eight samples of hematological malignancies that were scanned for BRAF p.V600E mutation using High resolution melting analysis (HRM) alone and combined HRM and COLD PCR. Results:When HRM was used alone for BRAF p.V600E mutation screening only 3 cases out of 38 were positive for the mutation, while when HRM was combined with COLD PCR 8 cases (21.1%) were positive for the mutationined HRM with COLD PCR compared to HRM alone and with Sanger sequencing

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