2.4 Detection of BRAF-V600E mutation in lesions and
cfDNA
We performed targeted DNA sequencing to detect mutations in LCH lesions,
using a customized genes panel that covers 229 genes involved in MAPK,
PI3K, or Jak/STAT, Epithelial-mesenchymal transition, cell cycle, cell
proliferation, apoptosis, cell adhesion/migration, and transcriptional
regulation (MyGenostics Inc.). We determined BRAF-V600E levels in plasma
cell-free (cf) DNA using QX200TM System Droplet Digital PCR (ddPCR)
System (Bio-Rad, Hercules, CA), as described
previously32.