A portable microfluidic platform for on-site detection of Aeromonas hydrophila in aquaculture water using...

Aeromonas hydrophila is a significant pathogenic bacterium in aquaculture, causing severe diseases in farmed animals and resulting in substantial economic losses. To enable rapid on-site detection, we developed a portable microfluidic platform that integrates multi-enzyme isothermal rapid amplification (MIRA) with automated fluid handling. The chip design incorporated a miniature vibration motor and ceramic heating pads to maintain precise temperature control and ensure reaction homogeneity throughout nucleic acid extraction and amplification. A miniature air pump drove the crude DNA extract through the chip in a single pipetting step for accurate quantification. The extracted DNA was subsequently transferred to a reaction chamber preloaded with MIRA reagents for isothermal amplification at 39 °C under real-time fluorescence monitoring. This platform detected A. hydrophila in water samples within 40 min, achieving a linear range from 1 × 10² to 1 × 10⁷ CFU·mL^-1 and a limit of detection (LOD) of 10 CFU·mL^-1. Notably, its modular design allows easy interchange of primers and probes, making the platform readily adaptable for detecting diverse bacterial pathogens in aquaculture.