therascreen FGFR RGQ RT-PCR Kit version 2

For qualitative detection of actionable alterations in the FGFR3 gene


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therascreen FGFR RGQ RT-PCR Kit version 2 (24)

Cat. No. / ID:  874721

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
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therascreen FGFR RGQ RT-PCR Kit version 2 (24)
Rotor-Gene Q MDx (US)
Rotor-Gene AssayManager v2.1
Rotor-Gene AssayManager v2.1 License
The therascreen FGFR RGQ RT-PCR Kit version 2 is intended for in vitro diagnostic use.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering


  • The only FDA-approved FGFR alteration assay on the market
  • 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

Product Details

The therascreen FGFR RGQ RT-PCR Kit version 2 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 version 2 is the only FDA-approved 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.


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 therascreen FGFR RGQ RT-PCR Kit and CTA (279/287 patients; 97.2%) 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 FGFR 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 85.2% (69/81) 75.9, 91.3
Negative percent agreement* 97.0% (192/198) 93.5, 98.6
Overall percent agreement 93.5% (261/279) 90.0, 96.1

* Prior chemotherapy information was not collected for CTA-negative patients. Therefore, 198 CTA-negative subjects included both chemo-relapsed/chemo-refractory patients and chemo-naïve patients.

The primary objective of the BLC2001 clinical trial was to evaluate the objective response rate (ORR) to treatment with BALVERSA in subjects with cases of UC in which FGFR alterations were detected. ORR was defined as complete response (CR) plus partial response (PR) by RECIST criteria, as assessed by blinded independent review committee (BIRC). The observed clinical benefit of treatment with BALVERSA in the subset of patients in which FGFR alterations were detected with both the therascreen FGFR RGQ RT-PCR Kit and the CTA (n = 69) was comparable to that observed in the full study population, as defined by the CTA only (n = 87). The ORR in patients in whom FGFR alterations were detected by both the therascreen FGFR RGQ RT-PCR Kit and the CTA, as assessed by BIRC, was 33.3% (95% CI: 23.4-45.1%).

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


The therascreen FGFR RGQ RT-PCR version 2 assay is based on the selective amplification of alterations in the FGFR genes using the Rotor-Gene Q MDx (US) 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 run controls and the sample results are valid and sufficient assay target amplification occurs below the predetermined cycle number cutoff threshold, the report will show the FGFR alteration(s) detected in each sample.


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 ready-to-use 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 (US) 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.

The therascreen FGFR RGQ RT-PCR Kit version 2 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 version 2 as an aid in the selection of such patients for treatment with BALVERSA (erdafitinib).


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 FDA-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


Kit Handbooks (4)
RNeasy DSP FFPE Kit 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 RNeasy DSP FFPE Kit
For use with Rotor-Gene Q MDx (US) instruments

October 2021
Confirmatory retest of positive results might be required

October 2022
Transition from Version 1 to Version 2 kit

October 2022
Brochures & Guides (1)
High-quality, nucleic acid purification for successful PCR and NGS experiments.
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.

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
Protocol Files (1)
Certificates of Analysis (1)


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
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