therascreen BRAF V600E RGQ PCR Kit

For qualitative detection of V600E mutations in the BRAF gene using real-time PCR

Products

The therascreen BRAF V600E RGQ PCR Kit is intended for in vitro diagnostic use.

QIAamp DSP DNA FFPE Tissue Kit (50)

Cat. No. / ID: 60404

For 50 DNA preps: QIAamp MinElute columns, Proteinase K, Buffers, and Collection Tubes (2 ml)

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Rotor-Gene AssayManager v2.1 License (1)

Cat. No. / ID: 9025620

Single license for the installation of Rotor-Gene AssayManager v2.1 software on one computer. For installation on multiple computers, each computer requires an individual computer-specific license key.Visit the Download Page for software download, installation and licensing instructions.

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Features

FDA-approved BRAF mutation companion diagnostic test
Reliable detection of V600E mutations in the BRAF gene
High sensitivity and specificity
Simple workflow from sample to insight
Automated data analysis using Rotor-Gene AssayManager for fast, easy result determination

Product Details

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.

Performance

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.

Principle

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.

Procedure

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.

Applications

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.

Resources

For the Directive 98/79/EC (IVDD) compliant kit (kit version 1)

QIAamp DSP DNA FFPE Tissue Handbook_V2_In Vitro Diagnostic use according to the Regulation (EU) 2017/746 on in vitro diagnostics medical devices

High-quality, nucleic acid purification for successful PCR and NGS experiments.

To download Rotor-Gene AssayManager v2.1.1 after purchase:

Rotor-Gene AssayManager v2.1.1 Software

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IMPORTANT

D24F10DEB7A1D60EE381E2AEA8789117D0980FCC

To purchase a license key for Rotor-Gene AssayManager v2.1.1:

Technical Support Form

IMPORTANT: The license file must always exist on the computer that has Rotor-Gene AssayManager v2.1.1 installed. The license file location should be protected and not subject to change.

QIAamp DSP DNA FFPE Tissue Performance Characteristics_V2_In Vitro Diagnostic use according to the Regulation (EU) 2017/746 on in vitro diagnostics medical devices

FAQ

Please refer to section 'Guidelines for effective multiplex assays' under "Important Notes" in the Rotor-Gene Multiplex PCR Handbook for suitable combinations of reporter dyes, or visit our Multiplex real-time PCR Resource site for additional information.

FAQ ID -9028

Reaction volumes suitable for use on the Rotor-Gene Q are:

Rotor-Disc 100: 30 µl x 100-wells, 10-25 µl reaction volume
Rotor-Disc 72: 0.1 ml x 72-wells, 15-25 µl reaction volume
Strip Tubes 0.1 ml: 0.1 ml x 72-wells, 10-30 µl reaction volume, strips of 4 tubes and caps
PCR Tubes 0.2 ml: 0.2 ml x 36-wells, 15-50 µl reaction volume, individual tubes with caps

FAQ ID -9030

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.

FAQ ID -9023

Please send the original OTV run file to QIAGEN Technical Service for further assistance.

FAQ ID -9022

For hardware related issues, please send the support package to QIAGEN Technical Service. Within the Rotor-gene Q software, click Help and select Send Support Email. In the new window, select the file that relates to the issue and email it to QIAGEN Technical Service.

FAQ ID - 9024

QIAGEN recommends the annual inspection service on Rotor-gene instruments, during which all application-critical modules of the Rotor-gene are inspected and tested and an OTV check is conducted. Performed tests and test results are documented in a GMP/GLP-compliant Report. In addition, the end users can perform the temperature calibration in the lab as needed using the Rotor-Disc OTV kit.

Note: The Rotor-Disc OTV kit requires the Rotor-Disc 72 Rotor and Rotor-Disc 72 locking ring.

FAQ ID -9025

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.).

FAQ ID -728

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.

FAQ ID -9027

Yes, all empty positions in the rotor of the Rotor-Gene Q have to be filled with empty tubes. This guarantees optimal temperature repartition in the Rotor-Gene Q chamber.

FAQ ID -9029