From lab to clinic: A portable and automated microfluidic nucleic acid detection instrument for rapid STI pathogen identification.

Sexually transmitted infections (STIs) remain a significant global public health challenge. The rapid and precise identification of these pathogens is crucial for effective diagnosis and treatment. Current polymerase chain reaction (PCR)-based assays, despite their widespread use, present several limitations in clinical settings, including high costs and technical complexity. To address these challenges, we developed a portable and automated microfluidic nucleic acid detection device for multiplex detection of three sexually transmitted pathogens via the fluorescence quantitative PCR (qPCR) assay. This integrated platform combines automated sample processing, multiplex qPCR amplification, and fluorescence detection within a single device. The entire closed-loop testing procedure, from automated nucleic acid extraction to result generation, requires only 40 min. The system demonstrates detection limits of 360 copies/ml for Neisseria gonorrhoeae (NG), 140 copies/ml for Ureaplasma urealyticum (UU), and 290 copies/ml for Chlamydia trachomatis (CT). Clinical validation revealed exceptional performance in multiplex detection of infected specimens, showing complete concordance (100 %) with standard diagnostic results while maintaining high specificity. These findings highlight the instrument's clinical potential and establish a new paradigm for automated, multiplex pathogen detection.