Single-Droplet Microsensor for Ultra-Short Circulating EFGR Mutation Detection in Lung Cancer Based on Multiplex...

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Oncology

Background:

• Liquid biopsy offers non-invasive cancer detection via circulating tumor DNA (ctDNA).
• A key challenge is detecting ultra-short ctDNA (usctDNA) with multiplexed assays using minimal sample volumes.
• Existing platforms often require PCR or Next-Generation Sequencing (NGS), limiting sensitivity for usctDNA.

Purpose of the Study:

• To develop a novel, non-PCR, non-NGS multiplexing platform for lung cancer-associated usctDNA detection.
• To address the unmet need for sensitive detection of multiple lung cancer mutations from minimal biofluid samples.
• To validate the performance of the developed microsensor technology for specific EGFR mutations.

Main Methods:

• Development of a single-droplet-based multiplexing microsensor technology named "Electric-Field-Induced Released and Measurement (EFIRM) Liquid Biopsy" (m-eLB).
• The m-eLB utilizes micro-electrodes coated with specific probes for multiplexed usctDNA detection within a single droplet.
• The prototype was tested for accuracy using synthetic nucleotides targeting three EGFR mutations (L858R, Exon 19 deletion, T790M).

Main Results:

• The m-eLB demonstrated high accuracy in detecting specific EGFR target sequences.
• Area Under the Curve (AUC) values for individual mutations were 0.98 (L858R), 0.94 (Exon 19 deletion), and 0.93 (T790M).
• The combined multiplexing assay for the three EGFR mutations achieved an AUC of 0.97.

Conclusions:

• The developed m-eLB platform shows significant promise for accurate, multiplexed detection of lung cancer-associated usctDNA.
• This technology addresses the need for sensitive detection of multiple mutations from minimal sample volumes without PCR or NGS.
• The EFIRM liquid biopsy approach offers a potential advancement in non-invasive lung cancer diagnostics.