Quantitative determination of target gene with electrical sensor.

Area of Science:

• Molecular Biology
• Biotechnology
• Analytical Chemistry

Background:

• Quantitative DNA detection is crucial for diagnostics and research.
• Existing methods often require complex instrumentation or are prone to contamination.
• Isothermal amplification techniques offer simplified DNA amplification without thermal cycling.

Purpose of the Study:

• To develop a novel electrical sensor for simultaneous DNA amplification and detection.
• To integrate loop-mediated isothermal amplification (LAMP) with capacitively coupled contactless conductivity detection (C4D).
• To establish a sensitive, rapid, and cost-effective method for quantitative gene determination.

Main Methods:

• Loop-mediated isothermal amplification (LAMP) was employed for DNA amplification.
• Capacitively coupled contactless conductivity detection (C4D) was utilized for real-time monitoring.
• The O26-wzy gene served as a model target for quantitative analysis.

Main Results:

• The integrated LAMP-C4D system enabled simultaneous amplification and detection of specific DNA sequences.
• Quantitative analysis was achieved by monitoring the threshold time of the LAMP reaction.
• A detection limit of 12.5 copy/μL was obtained within 30 minutes under optimal conditions.

Conclusions:

• The developed electrical sensor offers a robust, simple, and cost-effective solution for quantitative gene determination.
• Contactless conductivity detection eliminates carryover contamination risks, enhancing reliability.
• The method is suitable for both specialized laboratories and point-of-care settings due to its sensitivity and ease of use.