therascreen FGFR RGQ RT-PCR Kit (CE-IVD)

For qualitative detection of actionable alterations in the FGFR3 gene

Features

  • Detection of four point mutations and two fusions in the FGFR3 gene
  • High sensitivity and specificity
  • Simple workflow with next-day results
  • Automated data analysis using Rotor-Gene AssayManager v2.1 software

Products

The therascreen FGFR RGQ RT-PCR Kit is intended for in vitro diagnostic use.
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therascreen FGFR RGQ RT-PCR Kit (24)

Cat. No. / ID: 874711

For 24 reactions: Reverse Transcriptase, RT Buffer 1, RT Buffer 2, RT Primer Mix, Mutations-1 Reaction Mix, Mutations-2 Reaction Mix, Fusions-1 Reaction Mix, Fusions-2 Reaction Mix, Water for NTC, Water for sample dil., FGFR Positive Control, PC Diluent
HK$27,760.00
Add to cart
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RNeasy DSP FFPE Kit

Cat. No. / ID: 73604

For 50 RNA preps: RNeasy MinElute Spin Columns, Collection Tubes, Elution Tubes, Deparaffinization Solution, Proteinase K, RNase-Free DNase I, DNase Booster Buffer, RNase-Free Buffers and RNase-Free Water
HK$5,260.00
Add to cart
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Rotor-Gene Q MDx 5plex HRM Platform

Cat. No. / ID: 9002032

Real-time PCR cycler and High Resolution Melt analyzer with 5 channels (green, yellow, orange, red, crimson) plus HRM channel, laptop computer, software, accessories: includes 1-year warranty on parts and labor, installation and training not included
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Rotor-Gene Q MDx 5plex HRM System

Cat. No. / ID: 9002033

Real-time PCR cycler and High Resolution Melt analyzer with 5 channels (green, yellow, orange, red, crimson) plus HRM channel, laptop computer, software, accessories: includes 1-year warranty on parts and labor, installation and training
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Rotor-Gene AssayManager v2.1

Cat. No. / ID: 9024203

Software for routine testing in combination with Rotor-Gene Q and 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.

Product Details

The therascreen FGFR RGQ RT-PCR Kit is a qualitative in vitro diagnostic test for the detection of two point mutations in exon 7 [p.R248C (c.742C>T), p.S249C (c.746C>G)], two point mutations in exon 10 [p.G370C (c.1108G>T) and p.Y373C (c.1118A>G)] and two fusions (FGFR3-TACC3v1 and FGFR3-TACC3v3) in the FGFR3 gene.

The therascreen FGFR RGQ RT-PCR Kit is a companion diagnostic (CDx) test for the identification of patients with cases of urothelial cancer (UC) that harbor these actionable FGFR alterations, and for whom treatment with BALVERSA (erdafitinib) is indicated.

Performance

Concordance between the clinical trial assay (CTA) used to identify patients for recruitment to the Phase 2 clinical trial of BALVERSA (42756493-BLC2001) and the therascreen FGFR RGQ RT-PCR Kit (CDx) was evaluated in a bridging study (1).

Samples with valid results for both the therascreen FGFR RGQ RT-PCR Kit and CTA (292 samples) were analyzed to assess the positive percent agreement (PPA), negative percent agreement (NPA) and overall percent agreement (OPA), based on agreement between the two methods for overall FGFR gene alteration status (i.e., FGFR Alteration Detected or Alteration Not Detected). Results are shown in Table 1.

Table 1. therascreen FGFR RGQ RT-PCR Kit versus CTA (with CTA as reference method)

Measure of agreement Percent agreement % (N) Two-sided 95% CI
Positive percent agreement (PPA) 87.2% (82/94) 79.0, 92.5
Negative percent agreement (NPA) 97.0% (192/198) 93.5, 98.6
Overall percent agreement (OPA) 93.8% (274/292) 90.5, 96.1

The primary objective of the 42756493-BLC2001 study was to evaluate the objective response rate (objective response rate [ORR] = complete response [CR] + partial response [PR]) by RECIST criteria as assessed by investigator assessment. The ORR by Blinded Independent Review Committee (BIRC) was also determined. The observed clinical benefit in the subset of patients that were FGFR Alteration Detected with the therascreen FGFR RGQ RT-PCR Kit (n = 81) was comparable to that observed in the full study population (n = 99).

The therascreen FGFR RGQ RT-PCR Kit is therefore indicated for use as an aid in identifying patients with cases of urothelial cancer that harbor these alterations and are therefore eligible for treatment with BALVERSA (erdafinitib).

Principle

The therascreen FGFR RGQ RT-PCR Kit is based on the selective amplification of alterations in the FGFR3 gene using the Rotor-Gene Q MDx 5plex HRM instrument for sensitive and specific analysis. The assay exploits the qPCR oligonucleotide hydrolysis principle using TaqMan probes.

Allele-specific technology allows accurate and highly reproducible detection of alterations, based on the use of specific forward and reverse primers and probes; only a perfect match between the primers and probes with the target cDNA allows extension and amplification in the PCR reaction. Result reporting is fully automated. If both the positive and no template controls are valid and the sample internal controls are valid, the FGFR alteration status will be displayed in the software.

Procedure

The fast and simple workflow takes ~12 hours from Sample to Insight.

RNA is first prepared from formalin-fixed paraffin-embedded (FFPE) urothelial tumor samples using the RNeasy DSP FFPE Kit. Purified RNA is then reverse transcribed using Reverse Transcriptase to generate cDNA for real-time PCR analysis. Optimized reverse transcription reagents and PCR master mixes provided in the kit enable high-fidelity reverse transcription and sensitive real-time PCR on the Rotor-Gene Q MDx 5plex HRM instrument. Qualitative results are displayed in Rotor-Gene AssayManager software, informing the system operator if one or more of the four point mutations and two fusions in the FGFR3 gene detected by the kit are present in each sample.

The therascreen FGFR RGQ RT-PCR Kit is also designed to identify FGFR2 fusions FGFR2-BICC1 and FGFR2-CASP7 and FGFR3 fusion FGFR3-BAIAP2L1, because patients harboring these FGFR fusions were eligible for the BLC2001 clinical trial. However, the test is not clinically validated to detect these three fusions. Drug safety and efficacy has not been established for cases of UC harboring these fusions and no claims are made for the use of the therascreen FGFR RGQ RT-PCR Kit as an aid in the selection of such patients for treatment with BALVERSA (erdafitinib).

Applications

The therascreen FGFR RGQ RT-PCR Kit enables qualitative detection of four point mutations and two fusions of the FGFR3 gene for in vitro diagnostic use. It is the only approved CDx assay for the selection of patients with cases of urothelial cancer (UC) harboring actionable alterations, for whom treatment with BALVERSA (erdafitinib) is indicated.

 

1. Janssen Biotech, Inc. Data on file.

Supporting data and figures

Resources

Protocol Files (1)
Version 1.0.1
Brochures & Guides (1)
High-quality, nucleic acid purification for successful PCR and NGS experiments.
Kit Handbooks (3)
RNeasy FFPE DSP Kit
PDF (295KB)
For verification of thermal accuracy of Rotor-Gene real-time cyclers
For in vitro diagnostic use

Operating Software (3)
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.





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 Plug-in for use with Rotor-Gene AssayManager v2.1
Instrument User Manuals (4)
For use with Rotor-Gene AssayManager v2.1 Software
For installation and setup of Rotor-Gene AssayManager v2.1
Rotor-Gene Q MDx User Manual, revision R3, for Rotor-Gene Q MDx instruments using Rotor-Gene Q Software version 2.3.4
For use with Rotor-Gene Q Software version 2.3.4
Technical Information (2)
Software User Guides (1)
For use with Rotor-Gene AssayManager v2.1 Gamma Plug-in
Performance Data (1)