Detection of EGFR mutations at pM concentration in ten minutes using a microfluidic concentration and separation...

Epidermal growth factor receptor (EGFR) mutation detection is now commonly used in the management of cancer patients, particularly those diagnosed with non-small cell lung cancer. Molecular beacon-based sensing is direct and rapid, but its sensitivity is low. Conversely, high-sensitivity detection methodologies based on amplification are robust and sensitive but are limited by relatively require long turnaround times. In this study, we utilized a size-resolved, molecular beacon-based strategy for the rapid detection of EGFR genomic alterations, specifically exon 19 deletions and L858R point mutation. This technology combines a concentration and separation module, which allows us to successfully demonstrate the detection of deletions and point mutations of EGFR in five minutes with a mutant allele sensitivity of 10%. The use of a dual-color detection insures fast detection with a reduced risk of false positives. This work represents a first step toward the fast and specific detection of genetic mutations to improve the management of patients with hard-to-treat tumors.