Comparison of methodologies for the detection of BRAF mutations in bone marrow trephine specimens. Issue 6 (14th March 2019)

• Record Type: Journal Article
• Authors: Cardus, Beatrix; Colling, Richard; Hamblin, Angela; Soilleux, Elizabeth
• Is Part Of: Journal of clinical pathology. Volume 72:Issue 6(2019)
• Page Start: 406
• Page End: 411
• Publication Date: 2019-03-14
• Subjects: Hairy cell leukaemia -- bone marrow -- BRAF -- molecular pathology -- immunohistochemistry; Pathology -- Periodicals; Pathology, Molecular -- Periodicals; 616.0705
• DOI: 10.1136/jclinpath-2019-205734
• Languages: English
• ISSNs: 0021-9746

Abstract

Abstract : 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.