American Journal of Clinical Pathology · 2013 · 33 citations · 20 references
MedicineOncologyPathologyMolecular DiagnosticsMolecular OncologyNeoplasiaCancer Molecular PathologyDna SequencingMutation AnalysisNext-generation Sequencing In OncologyPolymerase Chain ReactionThyroid CancerBraf V600 MutationsBraf V600e MutationMutant DetectionNext-generation Sequencing-based DetectionNucleic Acid AmplificationPolymerase Chain Reaction (Pcr)-based DiagnosisPrognostic Factors AnalysisReal-time Polymerase Chain Reaction (Pcr)
We used pyrosequencing, peptide nucleic acid (PNA)-clamping polymerase chain reaction (PCR), and real-time PCR to detect the BRAF V600E mutation and to investigate the prognostic effect of the BRAF V600E mutation in paraffin block specimens from 100 patients diagnosed with papillary thyroid carcinoma. Positive rates of PNA-clamping PCR, real-time PCR, and pyrosequencing were 66%, 70%, and 68%, respectively. Pyrosequencing and PNA-clamping PCR detected mutant type in a 99:1 (wild-type: mutant) DNA concentration, and PNA-clamping PCR detected mutant type in a 99.5:0.5 DNA concentration. Clamping PCR showed higher κ value than real-time PCR (0.729 vs 0.626). The BRAF V600E mutation was associated with an advanced stage of cancer (P = .045) and was found to be associated with poor prognostic factors. This study suggests that pyrosequencing can be as sensitive as real-time PCR and that PNA-clamping PCR is a sensitive and reliable method to detect the BRAF V600E mutation.
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Mutations of the BRAF gene in human cancer
Helen Davies, Graham R. Bignell, Charles Cox et al. · Nature · 2002 · 10.7K citations
Chris Heid, Junko Stevens, Kenneth J. Livak et al. · Genome Research · 1996 · 6.2K citations · Full text