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therascreen PIK3CA RGQ PCR Kit

For qualitative detection of 11 mutations in the PIK3CA gene by real-time PCR

 

  • Reliable detection of clinically relevant mutations in the PIK3CA gene
  • High sensitivity and specificity
  • Results in less than two working days
  • Ready-to-use reagents and reaction mixes
  • Automated data analysis using Rotor-Gene AssayManager v2.1 software

The therascreen PIK3CA RGQ PCR Kit is a real-time qualitative in vitro diagnostic PCR test for the detection of 11 mutations in the phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA) gene using a sample of DNA extracted from either formalin-fixed, paraffin-embedded (FFPE) breast tumor tissue or K2EDTA anticoagulated blood plasma, taken from a patient with breast cancer.

The therascreen PIK3CA RGQ PCR Kit is the first companion diagnostic (CDx) test to be approved by the FDA to aid in the selection of patients with breast cancer who may be eligible for treatment with the a-selective PI3K-inhibitor, PIQRAY (alpelisib), in combination with the estrogen receptor antagonist, fulvestrant.

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Cat No./ID: 60404
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therascreen PIK3CA RGQ PCR Kit (24)
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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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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

Product Details

9
In vitro diagnostic medical device.
 
Performance

The SOLAR-1 study (CBYL719C2301) was a randomized, double-blinded, placebo-controlled, international, multicenter Phase III clinical trial that compared treatment with PIQRAY plus fulvestrant with placebo plus fulvestrant in men and postmenopausal women with hormone receptor-positive, HER2-negative advanced breast cancer that had progressed on or after aromatase inhibitor treatment. A total of 572 breast cancer patients were enrolled into two cohorts, with or without a PIK3CA mutation. Patients were randomized to receive PIQRAY 300 mg plus fulvestrant or placebo plus fulvestrant in a 1:1 ratio. The primary endpoint was progression-free survival (PFS) determined using RECIST v1.1 criteria, based on investigator assessment.

 

Results using FFPE tissue specimens

SOLAR-1 showed that in patients whose tumors harbored specific PIK3CA mutations, treatment with PIQRAY plus fulvestrant prolonged median PFS by a clinically meaningful 5.3 months (11 months overall) compared with treatment with placebo plus fulvestrant, and conferred an estimated 35% risk reduction in disease progression or death. Analysis of the CDx PIK3CA mutant-positive patient subset (172 patients) demonstrated that those receiving PIQRAY plus fulvestrant had an estimated 36% lower risk of disease progression or death (HR = 0.64; 95% CI: 0.48, 0.85) than patients receiving placebo plus fulvestrant.

By contrast, CDx PIK3CA mutant-negative patients (209) receiving PIQRAY plus fulvestrant had a shorter increase in median PFS of 0.8 months, with an estimated 27% lower risk of disease progression or death (HR = 0.73; 95% CI: 0.46, 1.17) than patients receiving placebo plus fulvestrant.

The importance of establishing PIK3CA mutation status when identifying patients for treatment with PIQRAY plus fulvestrant is therefore clear; only patients whose tumors harbor actionable PIK3CA mutations are likely to experience a clinically meaningful increase in PFS.

Results using plasma specimens

K2EDTA anticoagulated peripheral venous whole blood clinical plasma specimens collected from breast cancer patients randomized in SOLAR-1 prior to initiation of study treatment (baseline) were tested retrospectively with the therascreen PIK3CA RGQ PCR Kit to evaluate concordance between tissue and plasma results.

Of the 328 therascreen PIK3CA RGQ PCR Kit tissue-positive patients, 179 were therascreen PIK3CA RGQ PCR Kit plasma-positive. Of the 215 therascreen PIK3CA RGQ PCR Kit tissue-negative patients, 209 were therascreen PIK3CA RGQ PCR Kit plasma-negative. There were no invalid plasma results (Table 1).

Table 1. Correspondence table between therascreen PIK3CA RGQ PCR Kit tissue results and therascreen PIK3CA RGQ PCR Kit plasma results
     therascreen PIK3CA RGQ PCR Kit - tissue  
therascreen PIK3CA RGQ PCR Kit - plasma  Positive Negative Invalid  Total
Positive  179  6  1  186
Negative  149  209  5  363
Invalid  0  0  0  0
Total  328  215  6  549
 

Agreement (PPA, NPA and OPA) between the therascreen PIK3CA RGQ PCR Kit plasma and therascreen PIK3CA RGQ PCR Kit tissue results was calculated using the therascreen PIK3CA RGQ PCR Kit tissue results as reference (Table 2). The point estimates of PPA, NPA and OPA were 55%, 97% and 72%, respectively.

Table 2. Agreement between therascreen PIK3CA RGQ PCR Kit plasma results and therascreen PIK3CA RGQ PCR Kit tissue results using the therascreen PIK3CA RGQ PCR Kit tissue results as reference
Measure of agreement Percent agreement % (N) Two-sided 95% CI 
Positive percent agreement (PPA)  55% (179/328)  49.0%, 60.1%
Negative percent agreement (NPA)*  97% (209/215)  94.0%, 99.0%
Overall percent agreement (OPA)  72% (388/543)  67.5%, 75.2%
* 95% CI calculated using the Clopper-Pearson Exact method.

The therascreen PIK3CA RGQ PCR Kit therefore enables reliable selection of breast cancer patients with PIK3CA alterations who may be eligible for treatment with PIQRAY.

Principle

The therascreen PIK3CA RGQ PCR Kit is comprised of  six reaction mixes; one control reaction targeting exon 15 and five mutation-specific reactions utilized to detect 11 mutations in exons 7, 9 and 20 of the PIK3CA gene (Exon 7: C420R; Exon 9: E542K; E545A, E545D [1635G>T only], E545G, E545K, Q546E, Q546R; and Exon 20: H1047L, H1047R, H1047Y). Allele-specific technology allows accurate and highly reproducible detection of mutations; DNA is selectively amplified using ARMS primers, probes and PCR clamps, 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 above the cutoff, the report will show the PIK3CA alteration(s) detected in each sample.

Procedure

The simple and straightforward testing workflow begins with manual DNA extraction from either FFPE breast tumor tissue (using the QIAamp DSP DNA FFPE Tissue Kit) or from K2EDTA anticoagulated plasma (using the QIAamp DSP Circulating Nucleic Acid 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 one or more of the 11 mutations detected by the kit are present. The kit is a qualitative assay that can yield results in less than two working days.

Applications

The therascreen PIK3CA RGQ PCR Kit enables qualitative detection of 11 mutations in the PIK3CA gene for in vitro diagnostic use. It is an FDA-approved CDx assay to identify patients with cases of breast cancer harboring actionable alterations for whom treatment with PIQRAY (alpelisib) may be appropriate.

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Brochures & Guides (1)
High-quality, nucleic acid purification for successful PCR and NGS experiments.
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Selection Guides (1)
High-quality, nucleic acid purification for successful PCR and NGS experiments.
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Instrument User Manuals (4)
For use with Rotor-Gene Q Software version 2.3.4
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For use with Rotor-Gene Q Software version 2.3.4
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For use with Rotor-Gene AssayManager v2.1 Gamma MDx (US) Plug-in
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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.






Gamma MDx Plug-in for use with Rotor-Gene AssayManager v2.1 MDx (US)
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For use on the Rotor-Gene Q. Rotor-Gene Q software 2.3.5 is compatible with Windows 7 and Windows 10 operating systems
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