Langerhans cell histiocytosis (LCH) is a rare inflammatory myeloid neoplastic disease driven by activating mutations in the mitogen-activating protein kinase signalling pathway, including the BRAF V600E mutation and BRAF deletions (BRAFdel). Next-generation sequencing and whole exome sequencing (WES) are valuable and powerful approaches for BRAFdel identification, but these techniques are costly and time consuming. Pyrosequencing is an alternative method that has the potential to rapidly and reliably identify gene deletions. We developed a custom pyrosequencing assay to detect the exon-12 BRAFdel in 18 biopsies from adult patients with LCH, which were all genotyped in parallel using Sanger sequencing and WES. A BRAFdel was detected in 7/18 (39%), 6/18 (33%) and 3/18 (17%) LCH lesions using WES, pyrosequencing and Sanger, respectively, with good concordance between the WES and pyrosequencing results (Kappa-coefficient=0.88). Therefore, our pyrosequencing assay is reliable and useful for detecting BRAFdel, particularly in BRAF V600E-negative LCH lesions, for which targeted treatment is indicated.
- molecular biology
Statistics from Altmetric.com
FJ and AS are joint first authors.
Handling editor Runjan Chetty.
Contributors FJ and SM interpreted the data, drafted the manuscript and revised the manuscript. VM analysed and interpreted the histological findings. EB managed the clinical database. GL managed the sample acquisition and collected the clinical data. AS performed the molecular analysis. BL analysed the data. SM and AT designed the research, analysed the data, interpreted the data and revised the manuscript.
Funding The authors have not declared a specific grant for this research from any funding agency in the public, commercial or not-for-profit sectors.
Competing interests GL declares travel accommodations from Vitalaire and Chiesi. SM declares advisory roles for Roche and Biocartis. AT declares speaker fees from Chiesi and travel accommodations from Boehringer Ingelheim, Astrazeneca, and Vitalaire.
Provenance and peer review Not commissioned; internally peer reviewed.
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.