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.
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therascreen BRAF V600E RGQ PCR Kit (24)

Cat. No. / ID:  874821

For 24 reactions: Control Reaction Mix, V600E Reaction Mix, BRAF Positive Control, Taq DNA Polymerase, Water for NTC, Water for Dilution
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$2,871.00
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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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$360.00
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Rotor-Gene Q MDx System (US)

Cat. No. / ID:  9002036

Real-time PCR cycler with 6 channels (the red and HRM channels are not intended for use with FDA cleared or approved nucleic acid tests), laptop computer, software, accessories, 1-year warranty on parts and labor, installation and training
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Rotor-Gene Q MDx Platform (US)

Cat. No. / ID:  9002035

Real-time PCR cycler with 6 channels (the red and HRM channels are not intended for use with FDA cleared or approved nucleic acid tests), laptop computer, software, accessories, 1-year warranty on parts and labor
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Rotor-Gene AssayManager v2.1

Cat. No. / ID:  9024203-US

Software for routine testing in combination with Rotor-Gene Q MDx instruments. IMPORTANT: Does not include license for use. License key must be purchased separately. Visit the Download Page for download, installation and licensing instructions.
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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

Kit Handbooks (3)
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
For in vitro diagnostic use

Rx only (For prescription use only)

For use with QIAamp DSP DNA FFPE Tissue Kit

For use with Rotor-Gene Q MDx (US) instruments
Brochures & Guides (1)
High-quality, nucleic acid purification for successful PCR and NGS experiments.
Instrument User Manuals (3)
For use with Rotor-Gene Q Software version 2.3.4
For use with Rotor-Gene Q Software version 2.3.4
For use with Rotor-Gene AssayManager v2.1 Gamma MDx (US) Plug-in
Operating Software (3)
For use on the Rotor-Gene Q. Rotor-Gene Q software 2.3.5 is compatible with Windows 7 and Windows 10 operating systems
Gamma MDx Plug-in for use with Rotor-Gene AssayManager v2.1 MDx (US)
To download Rotor-Gene AssayManager v2.1.1 after purchase:
    Click the Rotor-Gene AssayManager v2.1.1 Software header. On the next window, click Continue. Follow the on-screen instructions to install Rotor-Gene AssayManager v2.1.1.
    Rotor-Gene AssayManager v2.1.1 and license must be purchased prior to use.    

    IMPORTANT: In order to perform and analyze runs, at least one Rotor-Gene AssayManager v2.1.1 plug-in needs to be installed. Refer to your assay kit handbook for more information.

    IMPORTANT: To allow checksum integrity verification, use the value D24F10DEB7A1D60EE381E2AEA8789117D0980FCC.

To purchase a license key for Rotor-Gene AssayManager v2.1.1:
    1. After purchasing Rotor-Gene AssayManager v2.1.1 software and licenses, click the "Download" button and follow the onscreen instructions to install. After successful installation of the Rotor-Gene AssayManager v2.1.1 and a plug-in, a “key file” will be generated. This "key file" will be attached to the "Technical Support Form" in Step 5, below.   
    2. Go to the Technical Support Form. Make sure to include all required information and your Rotor-Gene Q or Rotor-Gene Q MDx serial number when filling out the form.       
    3. For Question 2, "Type of Request", select “Other”.
    4. For Question 4, “Your inquiry”, enter "Rotor-Gene AssayManager license request” and include your purchase order number, from your order confirmation.
    5. Include the "key file" from Step 1 as an attachment.
    6. Submit the form. QIAGEN Technical Services will respond via email with a license file within 1–2 working days.    
    7. After receiving your computer-specific Rotor-Gene AssayManager v2.1.1 license file, archive the license file locally on your computer.

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.





Protocol Files (1)
BRAF US RGAM assay profile
Certificates of Analysis (1)

FAQ

Which-reporter-dyes-can-be-combined-for-use-in-multiplex-PCR-on-the-Rotor-Gene-Q-Cycler?
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
What reaction volume is suitable for use in the Rotor-Gene Q?

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
Why is no fluorescence signal detected in my run?

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
What should I do if the Rotor-Disc OTV run does not pass?

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

FAQ ID -9022
What kind of file is required for hardware-related trouble-shooting?
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
Is regular calibration needed with the Rotor-gene instrument?
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
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

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
Can I import a standard curve from a previous PCR run on the Rotor-Gene Q?

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
Must I fill blank positions with empty tubes when running sample numbers lower than the rotor capacity of the Rotor Gene Q?
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