Visual and modular detection of pathogen nucleic acids with enzyme-DNA molecular complexes

Area of Science:

• Biotechnology
• Molecular Biology
• Nanotechnology

Background:

• Rapid and accurate detection of pathogen nucleic acids is crucial for effective infection management.
• Existing methods can be time-consuming, require specialized equipment, or lack sensitivity.
• There is a need for versatile and accessible diagnostic tools for pathogen identification.

Purpose of the Study:

• To develop a novel, modular detection platform for rapid, visual identification of pathogen nucleic acids.
• To demonstrate the system's capabilities in detecting diverse pathogen targets and performing molecular typing.
• To evaluate the platform's performance in clinical samples.

Main Methods:

• Development of enzyme-assisted nanocomplexes (enVision) integrating enzyme-DNA nanostructures for recognition and signaling.
• Utilizing enzymatic cascades for rapid, naked-eye color readout (<2 hours, <10 amol sensitivity).
• Implementation on a configurable microfluidic platform for enhanced programmability and smartphone quantification.

Main Results:

• The enVision platform achieved rapid (<2 hours) and sensitive (<10 amol) detection of pathogen nucleic acids.
• The system demonstrated versatility in detecting diverse targets and performing computational analysis on a microfluidic platform.
• Application to patient endocervical samples showed improved inter-subtype differentiation and expanded intra-subtype coverage for infection molecular typing.

Conclusions:

• The enVision platform offers a fast, sensitive, and visually interpretable method for pathogen nucleic acid detection.
• Its programmability and smartphone compatibility make it a versatile tool for point-of-care diagnostics and molecular epidemiology.
• This technology enhances the ability to identify and differentiate infections, including previously undetectable ones.