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Detection of BRAF V600E activating mutation in papillary thyroid carcinoma using PCR with allele-specific fluorescent probe melting curve analysis Rowe LR; Bentz BG; Bentz JSJ Clin Pathol 2007[Nov]; 60 (11): 1211-5BACKGROUND: A single hotspot mutation at nucleotide 1799 of the BRAF gene has been identified as the most common genetic event in papillary thyroid carcinoma (PTC), with a prevalence of 29-83%. AIMS: To use a PCR assay to molecularly characterise the BRAF activating point mutation in a series of PTC and benign thyroid cases and correlate the mutation results with histological findings. METHODS: Formalin-fixed paraffin-embedded (FFPE) sections were evaluated for the BRAF V600E mutation using LightCycler PCR with allele-specific fluorescent probe melting curve analysis (LCPCR). RESULTS: 42 (37 PTC; 5 benign) surgical tissue samples were analysed for the BRAF V600E activating point mutation. Using LCPCR and direct DNA sequencing, the BRAF mutation was identified in 23/37 (62.2%) PTC FFPE samples. DNA sequencing results demonstrated confirmation of the mutation. CONCLUSIONS: Detection of BRAF-activating mutations in PTC suggests new approaches to management and treatment of this disease that may prove worthwhile. Identification of the BRAF V600E activating mutation in routine FFPE pathology samples by a rapid laboratory method such as LCPCR could have significant value.|*Point Mutation[MESH]|Alleles[MESH]|Base Sequence[MESH]|Carcinoma, Papillary/*genetics[MESH]|DNA Mutational Analysis/methods[MESH]|DNA, Neoplasm/genetics[MESH]|Fluorescent Dyes[MESH]|Humans[MESH]|Molecular Sequence Data[MESH]|Polymerase Chain Reaction/methods[MESH]|Proto-Oncogene Proteins B-raf/*genetics[MESH]|Retrospective Studies[MESH]|Thyroid Neoplasms/*genetics[MESH] |
