DiaCarta's QClamp® EGFR Codon Specific Mutation Detection kit is a polymerase chain reaction (PCR)-based assay that uses XNA probes to suppress amplification of WT-DNA, selectively amplifying mutant DNAs

The testing procedure involves four (4) simple steps:1. Isolation of DNA from tumor biopsies, paraffin-embedded sections (FFPE), fresh frozen tumors, or tumor cell lines2. Amplification of mutant all DNAs at specified codons within the KRAS gene 3. Detection of amplification signal using a real-time PCR instrument capable of SYBR green detection 4. Documentation and interpretation of results. This test can be completed in approximately 2-3 hours from DNA to test result. Limit of Detection <0.1% RUO, <1% CE-IVD. Includes WT and Positive Controls. 6 reactions per sample. Minimal sample input (5-10ng per reaction)

The epidermal growth factor receptor (EGFR) gene encodes a membrane protein that possesses an intracellular receptor tyrosine kinase and is expressed on the surface of epidermal cells. EGFR plays a central role in transmitting signals that promote cell growth and proliferation. Its tyrosine kinase (TK) domain activates several downstream effectors that lead to activation of the Ras-Raf-MAPK pathway. Overexpression and oncogenic mutations that constitutively activate the TK domain of EGFR have been found in various solid tumors.

The presence of mutations in the EGFR oncogene correlates with response to certain tyrosine kinase inhibitor (TKI) cancer therapies in patients with non-small cell lung cancer (NSCLC). Such mutations in the EGFR oncogene are present in the general population of patients with NSCLC at a frequency of approximately 10% in patients from the USA, Europe, or Australia and up to 30% in patients from Japan and Taiwan (1, 2, 9). Excessive activation of EGFR has been shown to be associated with advanced stages of cancer and a poor prognosis.

Due to its association with malignancies, epidermal growth factor receptor (EGFR) has become the target of an expanding class of anti-cancer therapies, such as GILOTRIF, GEFITINIB (Iressa) and ERLOTINIB (Tarceva), which are tyrosine kinase inhibitors (TKIs). The presence of the T790M ‘gatekeeper’ mutation correlates with resistance to tyrosine kinase inhibitor (TKI) therapy. Detection of somatic mutations in the EGFR gene provides a powerful strategy to predict the response of cancer patients to selected therapeutic regimens (e.g., tyrosine kinase inhibitors in lung cancer), in an effort to increase the survival rate of cancer patients receiving targeted therapies.

The assay identifies the presence of all mutations in and near EGFR Codon 719 (Exon 18), Ex19Deletions (Exon 19), Codon 790(Exon 20), Codon 858 and 861 (Exon 21). All mutations at a particular codon are detected, but the exact nature of the mutation is not specified.