PiezoFusion Enables Physically Driven Sample-to-Answer Nucleic Acid Testing from Complex Samples

Area of Science:

• Microfluidics
• Molecular Diagnostics
• Biotechnology

Background:

• Complex food matrices pose challenges for point-of-need nucleic acid testing due to difficulties in sample preparation.
• Existing methods often require multiple steps, specialized reagents, and are prone to contamination.

Purpose of the Study:

• To develop a compact, integrated microfluidic architecture for reagent-minimal nucleic acid testing from complex food samples.
• To demonstrate the efficacy of this system for rapid detection of Salmonella Typhimurium.

Main Methods:

• Development of PiezoFusion, a microfluidic system featuring dual-stage filtration, piezoelectric-driven reagent-free lysis, and magnetic-chain nucleic acid capture.
• Integration of sealed isothermal amplification and visual readout for a complete sample-to-answer workflow.
• Testing with Salmonella Typhimurium across seven food and water matrices, benchmarked against quantitative polymerase chain reaction (qPCR).

Main Results:

• PiezoFusion successfully processed heterogeneous samples into amplification-compatible nucleic acids within 60 minutes, without biochemical lysis reagents.
• The system demonstrated high accuracy in detecting Salmonella Typhimurium, with 86.96% sensitivity, 100% specificity, and 96.43% overall accuracy across various matrices.
• Consistent positive/negative classification was observed across all tested food and water-associated matrices.

Conclusions:

• PiezoFusion offers a reagent-minimal, integrated solution for sample-to-answer nucleic acid testing from complex food samples.
• The developed microfluidic architecture provides a practical framework for field-compatible molecular diagnostics.
• The modular design allows for adaptation to other nucleic acid targets with appropriate assay design and lysis optimization.