Human EGFR 29 Gene Mutation Detection Kit (Real-time PCR)

The Ultrassay EGFR Mutations Detection Kit is a sensitive and selective assay for the detection of the most informative somatic mutations in the epidermal growth factor receptor (EGFR) gene. It is used clinically in China to select NSCLC patients who are most likely to respond to Iressa (Gefitinib) or Tarceva (Erlotinib).

The EGFR assay tests for a total of 29 mutations in eight PCR tubes, including both activating and resistance mutations. The presence of activating mutations indicates predicted response to mutant EGFR-targeting drugs. If the tumor tissue harbors resistance mutations, it may respond poorly to therapies designed to inhibit EGFR signalling.

[Background]

The American Society for Clinical Oncology (ASCO) recommends that patients with NSCLC who are being considered for first-line therapy with an EGFR TKI (patients who have not previously received chemotherapy or an EGFR TKI) should have their tumor tested for EGFR mutations to determine whether an EGFR TKI or chemotherapy is the appropriate first-line therapy. (Keedy, V.L. et al. (2011) J. Clin. Oncol. 29(15), 2121).

The European Committee for Medicinal Products for Human Use (CHMP) recommended that EGFR mutation status be tested in patients with metastatic non-small cell lung cancer to predict sensitivity to Iressa. (EPAR summary for the public. EMEA/280173/2009. EMEA/H/C/1016).

[Principles]

The EGFR kit employs proprietary real-time PCR technology to detect 29 mutations in the EGFR gene. Target DNA is amplified with mutation-specific PCR primers, and the mutant amplicons are detected with a novel fluorescent probe. For high quality DNA, about 2 ng of purified DNA is added to each PCR tube. For DNA from FFPE tissue older than three years, 10 to 15 ng of DNA is recommended. The test can detect mutations at a sensitivity of 1% ( i.e., one mutant copy to 99 wild-type copies).

The EGFR mutation kit enables detection of the following mutations:

•T790M

• L858R

• L861Q

• S768I

• G719S，G719A and G719C

• 3 insertions in exon 20 (detects the presence of any of 3 insertions but does not distinguish between them)

• 19 deletions in exon 19 (detects the presence of any of the 19 deletions but does not distinguish between them).

[Applicable Equipment]

—ABI7500 Real-Time PCR System

—Stratagene MX3000P/3005P Real-Time PCR System

—BioRad CFX96 Real-Time PCR System

—Ultrassay XP96 Real-Time qPCR System

—Hongshi SLAN-96P Real-Time PCR System