Role of EGFR Mutations in Disease

The epidermal growth factor receptor (EGFR) plays a crucial role in regulating cell growth, survival, proliferation, and differentiation through its involvement in several intracellular signaling pathways. As a member of the receptor tyrosine kinase (RTK) family, EGFR is located on the surface of many epithelial cells and is activated when specific ligands bind to its extracellular domain.

In healthy cells, EGFR activity is tightly controlled. However, in various malignancies—including non-small cell lung cancer (NSCLC), anal carcinoma, and glioblastoma—this balance is disrupted. Overexpression, amplification, or activating EGFR mutations can lead to unchecked cellular growth, contributing to tumor development and progression.

Among the most clinically significant EGFR mutations are point mutations and deletions within exons 18–21. These alterations result in constitutive activation of the receptor, which in turn continuously stimulates downstream oncogenic signaling pathways such as RAS-RAF-MEK-ERK and PI3K-AKT.

Two well-documented EGFR mutations—T790M and C797S—are associated with resistance to EGFR tyrosine kinase inhibitors (TKIs). T790M, located in exon 20, is the most common resistance mutation after first- and second-generation TKI therapy. C797S has emerged as a mechanism of resistance to third-generation inhibitors like osimertinib. Detecting these mutations is critical in deciding when to escalate or adjust therapy in patients with EGFR-mutated cancers.

Early and precise detection of EGFR mutations not only supports accurate diagnosis but also enables personalized treatment strategies that may improve patient outcomes, particularly in advanced lung cancer.

About the TRUPCR® EGFR PCR Kit

The TRUPCR® EGFR Real-time PCR Kit is an in vitro diagnostic solution designed to qualitatively detect 32 EGFR mutations across exons 18 through 21. These include critical mutations such as exon 19 deletions, L858R in exon 21, and resistance-related variants like T790M and C797S.

Built on ARMS (Amplification Refractory Mutation System) PCR technology, the kit ensures highly selective amplification of mutant alleles even in a background of wild-type DNA. This specificity enables detection of low-frequency mutations with a sensitivity of down to 1% mutant DNA, making it suitable for both tumor tissue and liquid biopsy samples.

Detection is accomplished using dual-labeled fluorescent probes (FAM and HEX), ensuring high signal clarity and reliable interpretation. The inclusion of an internal extraction control enhances the integrity of each run by helping avoid false-negative results, particularly in degraded or FFPE samples.

Whether your lab is working with fresh-frozen, FFPE tissues, or plasma-derived ctDNA, the TRUPCR® EGFR PCR Kit delivers consistent, reliable mutation detection to aid in clinical decision-making.

Principle and Workflow

The TRUPCR® EGFR PCR Kit uses ARMS PCR to differentiate between mutant and wild-type EGFR alleles. ARMS primers are designed to bind only to specific mutation sites, allowing for precise amplification.

The basic workflow includes:

1. DNA extraction from tumor tissue or plasma samples
2. Real-time PCR amplification using mutation-specific ARMS primers
3. Detection of amplification through fluorescent probes (FAM for mutations, HEX for controls)
4. Data interpretation using standard qPCR analysis software

The kit’s compatibility with a wide range of real-time PCR platforms, including ABI, Bio-Rad, and Roche instruments, ensures broad usability across laboratories. Its user-friendly protocol makes it an ideal solution for clinical labs looking to streamline their molecular diagnostics workflow without compromising accuracy.

Clinical Application of EGFR PCR Testing

The clinical relevance of EGFR mutation testing is particularly significant in the context of NSCLC, where targeted EGFR therapies have become a cornerstone of personalized treatment. The TRUPCR® EGFR PCR Kit helps clinicians:

• Stratify patients based on mutation status for first-line EGFR-TKI therapies
• Identify acquired resistance mutations, like T790M or C797S, which indicate the need for therapy adjustment
• Predict therapeutic response, guiding the use of first-, second-, or third-generation inhibitors
• Improve outcomes through earlier, evidence-based treatment selection

As more targeted therapies are developed, comprehensive mutation profiling will remain essential. The TRUPCR® EGFR PCR Kit provides a reliable, cost-effective, and scalable solution for labs committed to delivering actionable diagnostic insights.