1. Custom pyrosequencing assay to detect short BRAF deletions in Langerhans cell histiocytic lesions
- Author
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Aurélie Sadoux, Fanélie Jouenne, Gwenaël Lorillon, Emmanuelle Bugnet, Véronique Meignin, Abdellatif Tazi, Baptiste Louveau, and Samia Mourah
- Subjects
0301 basic medicine ,Sanger sequencing ,Mutation ,Myeloid ,Langerhans cell ,General Medicine ,Biology ,medicine.disease_cause ,medicine.disease ,Molecular biology ,Pathology and Forensic Medicine ,03 medical and health sciences ,symbols.namesake ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,Langerhans cell histiocytosis ,030220 oncology & carcinogenesis ,symbols ,medicine ,Pyrosequencing ,Histiocyte ,Exome sequencing - Abstract
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 BRAFV600E 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 BRAFV600E-negative LCH lesions, for which targeted treatment is indicated.
- Published
- 2020
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