Wettable Porous Arrays with Structural Color Barcodes Integration for Multiplex Urinary Tract Infection Bacteria Screening

Urinary tract infection (UTI) is one of the most common bacterial infections in clinical practice and is a potentially life-threatening infection with high morbidity, recurrence, and mortality rates. High-sensitivity multiplexed biomarker detection is crucial for the early diagnosis and treatment of UTI. Digital nucleic acid amplification technologies (dNAATs) offer highly sensitive molecular characterization by isolating individual template molecules and amplifying them separately. However, multiplex analysis with dNAATs often requires parallel amplification, which is time-consuming and labor-intensive. Here, we present a novel multiplex detection method that integrates structural color barcodes into superhydrophilic porous arrays. This method enables the sensitive detection of multiple targets simultaneously in a highly efficient manner. The excellent hydrophilicity of the microwells allows the reaction solution to fill each well, ensuring random and complete isolation of the nucleic acid molecules. The amplification reactions occur independently in each unit, enabling the absolute quantification of nucleic acids. The structural color barcodes fixed in the microwells have stable reflection peaks and large surface areas, serving as encoding carriers based on their distinct reflection wavelengths. This enables the screening of multiplexed UTI bacteria to be performed quickly, accurately, and reproducibly. By using different probes on the color barcodes, we were able to detect single and multiple UTI pathogens with high sensitivity. The results demonstrate that the integrated porous array platform with structural color barcodes realized the highly sensitive detection and screening of UTI bacteria, which has great potential in multiplex biomolecule detection and early diagnosis of infections.