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

Complex food samples remain challenging inputs for point-of-need nucleic acid testing because clarification, cell disruption, and nucleic acid purification must be integrated into a compact, low-intervention, and contamination-resistant workflow. Here we report PiezoFusion, a physically driven microfluidic architecture that couples dual-stage filtration, reagent-free pore-confined lysis powered by a piezoelectric microporous pump, and online magnetic-chain interception for continuous-flow nucleic acid capture and purification, followed by sealed isothermal amplification and visual readout. This architecture converts heterogeneous raw samples to amplification-compatible nucleic acids within 60 min without biochemical lysis reagents. Using Salmonella Typhimurium as a model target, we evaluated seven representative food and water-associated matrices, with 84 total tests benchmarked against qPCR. PiezoFusion achieved 86.96% sensitivity, 100% specificity, 93.48% balanced accuracy, and 96.43% overall accuracy, while maintaining consistent positive/negative classification across matrices. By integrating matrix clarification, reagent-free lysis, structured magnetic capture, and sealed amplification within a single workflow, PiezoFusion completes a reagent-minimal route to sample-to-answer nucleic acid testing from complex food samples and provides a practical framework for field-compatible molecular diagnostics. This work establishes a Salmonella-oriented PiezoFusion workflow and provides a modular framework that may be adapted to other nucleic acid targets after target-specific assay design and lysis optimization.