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therascreen EGFR RGQ PCR Kit version 2

For sensitive detection of the most frequently occurring somatic mutations in the EGFR oncogene in less than 4 hours

  • Compliance with EU IVD Directive 98/79/EC
  • Clinically validated using samples from the LUX-Lung III trial
  • Sensitive, robust test that can detect low levels of mutant DNA
  • Ready-to-use system with simple workflow
  • Highly selective and sensitive ARMS and Scorpions PCR technologies
The therascreen EGFR RGQ PCR Kit version 2 is a molecular diagnostic kit for detection of exon 19 deletions, L858R, L861Q, G719X, S768I, exon 20 insertions, and the resistance mutation T790M in the EGFR gene using real-time PCR on the Rotor-Gene Q 5plex HRM instrument. The kit provides all necessary reagents, and is optimized for rapid and sensitive detection of a low percentage of mutant DNA in a background of wild-type genomic DNA. ARMS PCR technology combined with the Scorpions detection technology provides exceptionally high sensitivity and specificity for the detection of specific mutations.
Cat No./ID: 874111
therascreen EGFR RGQ PCR Kit version 2 (24)
For 24 reactions: 1 Control Assay, 7 Mutation Assays, Positive Control, Taq DNA Polymerase
Cat No./ID: 9023537
therascreen EGFR Assay Package CD
Software protocol package for use with the therascreen EGFR RGQ PCR Kit and Rotor-Gene Q 5plex HRM or Rotor-Gene Q MDx 5plex HRM in combination with Rotor-Gene Q Software version 2.3 and a 72-well rotor.
Note:
The Assay Package is also available for download.

The therascreen EGFR RGQ PCR Kit version 2 is intended for in vitro diagnostic use in Europe.


Principle

The therascreen EGFR RGQ PCR Kit version 2 enables detection of exon 19 deletions, L858R, L861Q, G719X, S768I, exon 20 insertions, and the resistance mutation T790M in the EGFR gene. The EGFR gene encodes the epidermal growth factor receptor (EGFR) protein. Mutations in the tyrosine kinase domain of the EGFR gene can enable tumor growth and progression. EGFR mutations are found in approximately 10% of non-small cell lung cancer incidences in the US and 35% in East Asia.

The therascreen EGFR RGQ PCR Kit version 2 enables detection of 29 somatic mutations in the EGFR gene including:

19 deletions in exon 19
T790M
L858R
L861Q
G719X
S768I
3 insertions in exon 20

The kit utilizes two technologies — ARMS (Amplification Refractory Mutation System) and Scorpions — for detection of mutations using real-time PCR on the Rotor-Gene Q HRM 5plex instrument.

ARMS

Allele- or mutation-specific amplification is achieved by ARMS. Taq DNA polymerase is effective at distinguishing between a match and a mismatch at the 3' end of a PCR primer. Specific mutated sequences are selectively amplified, even in samples where the majority of the sequences do not carry the mutation. When the primer is fully matched, the amplification proceeds with full efficiency. When the 3' base is mismatched, only low-level background amplification occurs.

Scorpions

Detection of amplification is performed using Scorpions. Scorpions are bifunctional molecules containing a PCR primer covalently linked to a probe. The fluorophore in this probe interacts with a quencher, also incorporated into the probe, that reduces fluorescence. When the probe binds to the amplicon during PCR, the fluorophore and quencher become separated. This leads to an increase in fluorescence from the reaction tube.

Procedure
The therascreen EGFR RGQ PCR Kit version 2 uses a two-step procedure. The first step is performance of the control assay to assess the total DNA in a sample. The second step is to complete the mutation assay for the presence or absence of mutated DNA.
Applications

The therascreen EGFR RGQ PCR Kit version 2 enables detection of 29 somatic mutations in the EGFR gene. These 29 mutations correlate with responsiveness to tyrosine kinase inhibitor therapies, such as gefitinib and afatinib, for patients with non-small cell lung cancer. Some have a sensitizing effect and some are linked to resistance. The therascreen EGFR RGQ PCR Kit version 2 is compliant with EU IVD Directive 98/79/EC, enabling detection of mutations in the EGFR gene for in vitro diagnostics.

The therascreen EGFR RGQ PCR Kit version 2 enables detection of 29 somatic mutations in the EGFR gene including:

T790M, G719X, S768I
L858R, L861Q
3 insertions in exon 20
19 deletions in exon 19

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Handbooks & Protocols (2)

Important information for use with the therascreen EGFR RGQ PCR Kit with catalog number 874111 only. Please check your kit box label.


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Kit Handbooks (2)

Important information for use with the therascreen EGFR RGQ PCR Kit with catalog number 874111 only. Please check your kit box label.


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