Cat. No. / ID: 874821
Cat. No. / ID: 60404
Cat. No. / ID: 9002036
Cat. No. / ID: 9002035
Cat. No. / ID: 9024203-US
Cat. No. / ID: 9025620
The therascreen BRAF V600E RGQ PCR Kit is a qualitative in vitro diagnostic test for the detection of V600E mutations in the BRAF gene. The test analyzes DNA extracted from formalin-fixed, paraffin-embedded (FFPE) tumor tissue, taken from a patient with colorectal cancer (CRC). It is intended to aid clinicians in identifying metastatic colorectal cancer (mCRC) patients eligible for treatment with BRAFTOVI (encorafenib) in combination with cetuximab.
The clinical performance of the kit was determined in the BEACON CRC Study. This was a three-arm, multicenter, randomized, open-label, Phase 3 study of encorafenib + cetuximab plus (triplet arm) or minus (doublet arm) binimetinib versus irinotecan/cetuximab or infusional 5-fluorouracil/folinic acid/irinotecan/cetuximab (control arm) in patients with BRAF V600E mutant metastatic CRC.
The study comprised 665 patients (224 triplet arm; 220 doublet arm; 221 control arm). Study endpoints included overall survival (OS) and overall response rate (ORR) by BICR (Blinded Independent Central Review) per Response Evaluation Criteria in Solid Tumors (RECIST v1.1).
The study demonstrated a statistically significant clinical improvement in OS and confirmed ORR by BICR for the doublet arm versus the control arm, with a 40% reduction in risk of death observed with the doublet arm compared to the control arm (HR 0.60, 95% CI: 0.45, 0.79).
Therefore, there is a clear clinical benefit to determining BRAF mutation status when determining patient eligibility for treatment with encorafenib + cetuximab.
The therascreen BRAF V600E RGQ PCR Kit is comprised of one mutation and one control reaction mix utilized to detect the V600E mutation in the BRAF gene. Allele-specific technology allows accurate and highly reproducible detection of mutations; DNA is selectively amplified using ARMS primers and Scorpions probes, with sensitive signal detection using the Rotor-Gene Q MDx (US) instrument. Result reporting is fully automated. If both the run controls and the sample results are valid and target assay amplification is below the preset cutoff, the report will show the V600E mutation as detected in the sample.
The simple and easy testing workflow begins with manual DNA extraction from mCRC tumor tissue using the QIAamp DSP DNA FFPE Tissue Kit, followed by sensitive real-time PCR on the Rotor-Gene Q MDx (US) instrument. Rotor-Gene AssayManager software rapidly and accurately determines mutations and reports results, informing the system operator if the V600E mutation is present in the sample.
The therascreen BRAF V600E RGQ PCR Kit enables qualitative detection of V600E mutations in the BRAF gene for in vitro diagnostic use. It is an FDA-approved CDx assay to identify patients with cases of metastatic colorectal cancer for whom treatment with BRAFTOVI (encorafenib) in combination with cetuximab may be appropriate.
Reaction volumes suitable for use on the Rotor-Gene Q are:
Please make data are collected in the appropriate fluorescent channel. Also check the gain is optimized.
If the issue persists, please send the original run file with extension .rex to QIAGEN Technical Service for further assistance.
Please send the original OTV run file to QIAGEN Technical Service for further assistance.
Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.
Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).
Use of endogenous control genes corrects for variation in RNA content, variation in reverse-transcription efficiency, possible RNA degradation or presence of inhibitors in the RNA sample, variation in nucleic acid recover, and differences in sample handling. The endogenous control gene ought to have consistent expression levels between samples and among treatment conditions, and ideally has a similar expression level to that of the genes of interest. Genes commonly used as references can be found at the QuantiTect Primer Assays as endogenous controls.