ipsogen CALR RGQ PCR Kit CE

For broad, sensitive and reliable CALR mutation detection to improve patient diagnosis

Products

The ipsogen CALR RGQ PCR Kit is intended for in vitro diagnostic use. References: Tefferi, A. and Pardanani, A. (2014) CALR mutations and a new diagnostic algorithm for MPN. Nat. Rev. Clin. Oncol. 11:125. Arber, D.A. et al. (2016) The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood 127:2391. Klampfl T. et al. (2013) Somatic mutations of calreticulin in myeloproliferative neoplasms. N Engl J Med. 369(25):2379. Nangalia J. et al. (2013) Somatic CALR mutations in myeloproliferative neoplasms with nonmutated JAK2. N Engl J Med. 369(25):2391. Rumi, E., et al. (2014) Clinical effect of driver mutations of JAK2, CALR, or MPL in primary myelofibrosis. Blood 124:1062.
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ipsogen CALR RGQ PCR Kit (24) CE

Cat. No. / ID:  674023

For 24 reactions: Wild-type CALR Control, Mutant CALR Control, CALR TYPE1 Reaction Mix, CALR TYPE2 Reaction Mix, CALR CLAMP1 Reaction Mix, CALR CLAMP2 Reaction Mix, CALR CLAMP3 Reaction Mix, CALR CLAMP4 Reaction Mix, CALR CLAMP5 Reaction Mix, Taq DNA polymerase, TE buffer for dilution and NTC
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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.
For up-to-date licensing information or Rotor-Gene Q instrument specific disclaimers, refer to the corresponding Rotor-Gene Q instrument user manual

Features

  • Clinically reliable detection of CALR mutations and identification of Types 1 & 2, in a single run
  • Less effort with only one instrument and one real-time PCR step, for multiple clinical results in less than 1 working day
  • Analyze JAK2 V617F and CALR from the same patient sample
  • All-in-one kit with all required components
  • Facilitate ISO 15189 accreditation and routine implementation
  • Follow diagnostic algorithm testing (1) and WHO 2016 guidelines (2) by testing clinically relevant MPN biomarkers

Product Details

The ipsogen CALR RGQ PCR Kit is a ready-to-use CE-IVD kit intended for the detection of CALR mutations in genomic DNA from subjects suspected of myeloproliferative neoplasms (MPN). The ipsogen CALR RGQ PCR Kit also enables the identification of the two major CALR mutations (Type 1 and Type 2), and is to be used with the QIAGEN Rotor-Gene Q MDx 5Plex HRM Platform. The kit is intended for use by professionals, such as technicians and physicians, who are trained in molecular biology techniques.

Performance

To ensure sensitivity and accuracy, the ipsogen CALR RGQ PCR Kit has been optimized to perform seven separate PCR amplification reactions in a single run for the identification of the two major CALR mutations (Type 1 and Type 2) and the detection of additional minor variants in genomic DNA extracted from human peripheral whole blood. As shown in the figure entitled ' Results generated by the ipsogen CALR RGQ PCR Kit ', the kit identifies CALR mutations Type 1 and Type 2, and detects additional mutations in the CALR exon 9 region (c.1091_1162).

The limit of detection (LoD) of the kit was determined to be 0.60% for the Type 1 CALR mutation and 0.08% for the Type 2 CALR mutation, respectively. The clinical performance was validated using 227 patient samples and showed sensitivity of 100% and specificity of 97.7% at an overall concordance of 98.7% compared to sequencing-based reference methods. >

See figures

Principle

The ipsogen CALR RGQ PCR Kit is a real-time PCR test for the qualitative detection of somatic mutations in the region c.1091_1162 (cDNA annotation) of exon 9 in the CALR gene (GenBank Accession Number CR457070) (3, 4), and also enables the identification of the two major CALR mutations (Type 1 and Type 2). The kit provides reagents to perform seven separate PCR amplification reactions in the same run for the identification of CALR mutations Type 1 and Type 2, and the detection of additional minor variants in genomic DNA (gDNA) extracted from human peripheral whole blood. The turnaround time to execute all tasks, from gDNA extraction (using manual or automated extraction) to data analysis, is less than one working day.

To validate and control the PCR reaction in the presence of human gDNA template, each CALR reaction mix includes primers and probe to amplify an endogenous sequence of the ABL1 housekeeping gene. This control sequence is amplified through a multiplex PCR reaction in all CALR mutant and wild-type DNA. All required components are included in the run to provide accurate and sensitive detection of CALR mutations.

Results analysis and interpretation is automated and simplified with the Rotor-Gene AssayManager v2.1 software.

Procedure

The ipsogen CALR RGQ PCR Kit allows detection of CALR mutations and specific identification of Types 1 and 2 on Rotor-Gene Q MDx 5plex HRM. Genomic DNA is first extracted from human peripheral whole blood using the QIAamp DSP Blood Mini Kit (manual) or QIAsymphony SP-automated solution, followed by determination of quality and quantity, and gDNA amplification by real-time PCR. Results interpretation is automated using the Rotor Gene AssayManager v2.1 software. The kit provides all required reagents to perform 7 separate qPCR amplification reactions in the same run for detection of CALR mutations and identification of Types 1 and 2 in genomic DNA extracted from human peripheral whole blood. Simply start the reaction on the Rotor-Gene Q MDx 5plex HRM using the optimized protocols as described in the kit handbook.

Applications

The ipsogen CALR RGQ PCR Kit for in vitro diagnostic use enables sensitive and reliable detection of CALR exon 9 mutations, as well as identification of the two major CALR mutations (Type 1 and Type 2). Detection of CALR mutations is now part of the reference World Health Organization (WHO) 2016 criteria for the diagnosis of myeloproliferative neoplasms (MPN) (2), and presence of this mutation is a major criterion for diagnostic confirmation. The prognostic significance of CALR mutations have also been described (5).

Supporting data and figures

Resources

Operating Software (2)
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 Plug-in for use with Rotor-Gene AssayManager v2.1
Software User Guides (1)
For use with Rotor-Gene AssayManager v2.1 Gamma Plug-in
Instrument User Manuals (2)
For use with Rotor-Gene AssayManager v2.1 Software
For installation and setup of Rotor-Gene AssayManager v2.1
Safety Data Sheets (1)
Kit Handbooks (1)
Protocol Files (1)
Certificates of Analysis (1)