therascreen BRAF Assay Package CE

This CE-marked IVD from QIAGEN is a ready-to-use kit designed for the qualitative measurement of somatic mutations in the BRAF oncogene, using polymerase chain reaction (PCR) on the Rotor-Gene Q MDx 5plex HRM instrument. The BRAF gene encodes the kinase V-raf murine sarcoma viral oncogene homolog B1, a proto-oncogene that acts downstream of EGFR. The assay enables qualitative detection from genomic DNA extracted from formalin-fixed paraffin-embedded (FFPE) samples and is validated for molecular diagnostic workflows.

Five assays are supplied in the kit: one control assay (Control Reaction Mix; CTRL) and four mutation assays (V600E/Ec, V600D, V600K, V600R). The V600E/Ec assay detects both V600E and V600Ec mutations but does not distinguish between them. All reaction mixes are duplex and contain reagents to detect targets labeled with FAM and an internal control labeled with HEX. The internal control assay monitors the presence of inhibitors that may lead to false negative results, while the control assay amplifies a region of exon 3 of the BRAF gene.

Detection relies on two combined technologies: ARMS (Amplification Refractory Mutation System) for allele- or mutation-specific amplification, and Scorpions for amplification detection. Scorpions are bifunctional molecules containing a PCR primer covalently linked to a fluorescently labeled probe; when the probe binds to the amplicon during PCR, the fluorophore and quencher become separated, producing a measurable increase in fluorescence. The kit is supplied with positive and negative controls and complete instructions for use.