AIMS: BRAF V600E detection assists in the diagnosis of hairy cell leukaemia (HCL); however, testing practices vary. We evaluated the clinical utility of 5 BRAF mutation testing strategies for use on bone marrow trephines (BMT). METHODS: 11 HCL, 5 HCL 'mimic', 2 treated HCL and 10 normal BMT specimens were tested for mutant BRAF, comparing Sanger sequencing, pyrosequencing, amplicon-based next generation sequencing (NGS), automated (Idylla) PCR and immunohistochemistry (IHC). RESULTS: PCR and IHC were cheaper and identified V600E in 100 % of HCL cases. Pyrosequencing detected the mutation in 91%, NGS in 55% of cases and Sanger sequencing in 27%. All assays gave wild-type BRAF results in HCL mimics and normal BMT samples. CONCLUSIONS: PCR and IHC were most sensitive and cost-effective, but these have limited scope for multiplexing and are likely to be replaced by NGS gene panels or whole genome sequencing in the medium to long term.
Journal article
2019-06-01T00:00:00+00:00
72
406 - 411
5
BRAF, Hairy cell leukaemia, bone marrow, immunohistochemistry, molecular pathology, Automation, Laboratory, Biomarkers, Tumor, Biopsy, Bone Marrow, Bone Marrow Examination, Cost-Benefit Analysis, DNA Mutational Analysis, Health Care Costs, High-Throughput Nucleotide Sequencing, Humans, Immunohistochemistry, Leukemia, Hairy Cell, Mutation, Predictive Value of Tests, Proto-Oncogene Proteins B-raf, Real-Time Polymerase Chain Reaction, Reproducibility of Results