One-Click Total Bacterial Quantification: An All-in-One Automatic Microfluidic Platform for Simple Liquid Food Samples

Food safety detection is essential for disease prevention and public health protection. However, current bacterial detection technologies generally face challenges including complex operation, long detection cycles, and large sample requirements. While adenosine triphosphate (ATP) bioluminescence technology offers rapid, sensitive microbial quantification, traditional processes remain inefficient. To address these challenges, this study developed a fully automated integrated ATP bioluminescence bacterial quantification platform. The system integrates a microfluidic chip, automatic sample injection module, and real-time luminescence analysis photoelectric module, achieving an end-to-end workflow of mixing, on-chip reaction, analysis, and cleaning, eliminating manual intervention and improving convenience and efficiency. Chip’s advanced structures ensure high mixing efficiency of 98.9%, significantly reducing required sample volume and cost. With detection time under 10 min, experimental validation demonstrated excellent interbatch consistency and repeatability (RSD $ \lt {10}\% $ ), ATP dynamic detection range of 10 pM to $ {1}~\mu \text {M} $ ( $ {R}^{{2}} = {0.9995} $ ), and detection limit of 4.86 pM. For Vibrio parahaemolyticus, the system achieved a detection range of $ {10}^{{2}} $ – $ {10}^{{8}}~\text {CFU/mL} $ ( $ {R}^{{2}} = {0.996} $ ). In spiked Coca-Cola samples, the system maintained excellent anti-interference ability and accuracy ( $ {R}^{{2}} = {0.941} $ ), with standard curve slopes differing by only 11.3% from PBS buffer. This portable platform with high sensitivity, stability, and repeatability is suitable for field testing of water and simple liquid foods. In the future, by integrating specific bacterial capture technology and sample pretreatment modules, the system is expected to extend to more complex matrix sample detection, further enhancing its application value in food safety.