Development and validation of a real-time quantification assay to detect and monitor BRAFV600E mutations in hairy cell leukemia

作者：Schnittger Susanne^*; Bacher Ulrike; Haferlach Torsten; Wendland Nicole; Ulke Madlen; Dicker Frank; Grossmann Vera; Haferlach Claudia; Kern Wolfgang

来源：Blood, 2012, 119(13): 3151-3154.

DOI：10.1182/blood-2011-10-383323

The BRAFV600E mutation was recently detected in hairy cell leukemia (HCL) by whole exome sequencing. To make use of this new marker for diagnosis and follow-up of HCL, we developed a BRAFV600Emut-specific quantitative real-time PCR assay and validated it in 117 HCL patients and 102 non-HCL/BRAFwt patients. The cut-off level to discriminate BRAFV600E-positive/-negative cases was set at 0.023% BRAFV600E/BRAFwt. A total of 115 of 117 HCL (98.3%) demonstrated percentage BRAFV600E/BRAFwt above the cut-off (mean, 29.6 +/- 41.1). The remaining 2 of 117 HCL with lower percentage BRAFV600E/BRAFwt ratios were also BRAFwt by deep-sequencing technology. Sixteen HCL-variant patients showed percentage BRAFV600E/BRAFwt values corresponding to %26quot;non-HCL.%26quot; Follow-up studies in 19 HCL cases demonstrated a decrease of percentage BRAFV600E/BRAFwt during therapy. The log-reductions as determined by RT-PCR and immunophenotyping correlated significantly (P %26lt; .001). In conclusion, we confirmed BRAFmut as a useful marker in HCL, its absence in HCL variant, and developed an RT-PCR-based assay to monitor minimal residual disease in HCL. (Blood. 2012;119(13):3151-3154)

• 出版日期2012-3-29