Rapid detection and quantitation of BRAF mutations in hairy cell leukemia using a sensitive pyrosequencing assay.
BRAF protooncogene is an important mediator of cell proliferation and survival signals. BRAF p.V600E mutation was recently described as a molecular marker of hairy cell leukemia (HCL). We developed and validated a pyrosequencing-based approach that covers BRAF mutational hotspots in exons 11 (codon 468) and 15 (codons 595 to 600). The assay detects BRAF mutations at an analytical sensitivity of 5%. We screened 16 unenriched archived bone marrow aspirate samples from patients with a diagnosis of HCL (n = 12) and hairy cell leukemia-variant (HCL-v) (n = 4) using pyrosequencing. BRAF p.V600E mutation was present in all HCL cases and absent in all HCL-v. Our data support the recent finding that BRAF p.V600E mutation is universally present in HCL. Moreover, our pyrosequencing-based assay provides a convenient, rapid, sensitive, and quantitative tool for the detection of BRAF p.V600E mutations in HCL for clinical diagnostic testing.
Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Naomi St. Facility (NAO1.053a), 1515 Holcombe Blvd, Unit 0149, Houston, TX 77030, USA.
Shalini
S
Verma
Wesley O
WO
Greaves
Farhad
F
Ravandi
Neelima
N
Reddy
Carlos E
CE
Bueso-Ramos
Susan
S
O'Brien
Deborah A
DA
Thomas
Hagop
H
Kantarjian
L Jeffrey
LJ
Medeiros
Rajyalakshmi
R
Luthra
Keyur P
KP
Patel
D000328:Adult; D000368:Aged; D004252:DNA Mutational Analysis / Q000379:methods*; D005091:Exons; D005260:Female; D006801:Humans; D007943:Leukemia, Hairy Cell / Q000175:diagnosis* / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009154:Mutation; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*
C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf