TY - JOUR T1 - Comparison of methodologies for the detection of <em>BRAF</em> mutations in bone marrow trephine specimens JF - Journal of Clinical Pathology JO - J Clin Pathol SP - 406 LP - 411 DO - 10.1136/jclinpath-2019-205734 VL - 72 IS - 6 AU - Beatrix Cardus AU - Richard Colling AU - Angela Hamblin AU - Elizabeth Soilleux Y1 - 2019/06/01 UR - http://jcp.bmj.com/content/72/6/406.abstract N2 - 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. ER -