Ultra-Fast Single-Nucleotide-Variation Detection Enabled by Argonaute-Mediated Transistor Platform

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Biology

Background:

• Current single-nucleotide variation (SNV) detection methods often require lengthy procedures like nucleic acid amplification or sequencing, limiting rapid diagnostics.
• A significant trade-off exists between the sensitivity and speed of existing SNV detection techniques, particularly for low-abundance samples.

Purpose of the Study:

• To develop a rapid, sensitive, and specific method for "test-and-go" SNV detection.
• To overcome the limitations of conventional SNV detection by eliminating the need for sample preparation steps like nucleic acid extraction, reverse transcription, and amplification.

Main Methods:

• Development of a graphene field-effect transistor (GFET) platform integrated with Argonaute protein.
• Utilizing the Argonaute protein to create a nanoscale binding channel for preorganizing DNA probes, thereby accelerating target binding.
• Employing the GFET-Argonaute system for direct detection of SNVs in unamplified nucleic acid samples, including microRNA, circulating tumor DNA, and cDNA.

Main Results:

• The developed GFET platform enables SNV detection with single-nucleotide resolution.
• The system successfully identifies SNVs in the seed region of various nucleic acid targets, including tumor-associated microRNA, circulating tumor DNA, virus RNA, and reverse transcribed cDNA.
• An integrated microchip simultaneously detected multiple SNVs within 5 minutes, achieving results comparable to sequencing.

Conclusions:

• The Argonaute-mediated GFET platform offers a groundbreaking "test-and-go" solution for rapid and sensitive SNV detection.
• This approach significantly reduces detection time and complexity, paving the way for fasterPoint-of-care diagnostics.
• The technology demonstrates broad applicability for detecting SNVs in diverse biological samples without prior nucleic acid manipulation.