Copper: DNA extraction and solid phase detection agent for all-in-one molecular diagnostic device coupled with...

Area of Science:

• Biotechnology
• Microfluidics
• Analytical Chemistry

Background:

• Current pathogen detection methods often require complex laboratory equipment and trained personnel, limiting their use in resource-limited settings.
• Developing portable, equipment-free diagnostic tools is crucial for decentralized testing and rapid response to outbreaks.

Purpose of the Study:

• To develop an integrated, all-in-one poly(methyl methacrylate) (PMMA) device for on-site pathogen detection.
• To combine novel DNA extraction and detection methods with loop-mediated isothermal amplification (LAMP) for simplified molecular diagnostics.

Main Methods:

• Utilized a Fenton-like reaction (CuSO4/H2O2) for room-temperature DNA extraction via bacterial membrane disruption.
• Employed loop-mediated isothermal amplification (LAMP) for DNA amplification.
• Developed a visual detection method using copper tape corrosion, triggered by the acidic environment produced during LAMP with target DNA.

Main Results:

• Successfully detected Enterococcus faecium down to 4.67 × 10^2 CFU/mL within 90 minutes.
• Demonstrated successful detection of E. faecium spiked in milk samples.
• The device integrates DNA extraction, amplification, and naked-eye detection in a single unit.

Conclusions:

• The developed all-in-one PMMA device offers a promising, equipment-free solution for decentralized pathogen detection.
• The integration of novel DNA extraction, LAMP, and visual detection provides a portable and cost-effective point-of-care testing platform.
• This technology is particularly suitable for resource-limited environments requiring rapid and accessible diagnostics.