Argonaute Integrated with Nanoparticle-Based Biosensor for Portable, Visual, and Rapid Detection of Neisseria gonorrhoeae in Clinical Application

Sexually transmitted gonorrhea infection is a key worldwide public health issue, especially in low- and middle-income areas. The quest for ultrasensitive, highly specific, rapid, and accessible diagnostics for Neisseria gonorrhoeae is a critical component of global strategies to combat and treat gonorrhea. Herein, we first reported a PfAgo-N. gonorrhoeae diagnostic platform by integrating PfAgo-mediated detection with LAMP-based preamplification, and the results were analyzed via gold nanoparticle-based lateral flow biosensor (AuNP-LFB), real-time fluorescence intensity (RFI), or blue light. Our PfAgo-N. gonorrhoeae assay demonstrated a limit of detection (LoD) as low as five copies/test and presented no cross-reactivity with other tested pathogens. The detection procedure, comprising nucleic acid isolation (10 min), LAMP amplification (30 min), PfAgo detection (10 min), and AuNP-LFB visual readout (<2 min), can be completed within 55 min and does not require any specialized instrumentation. The clinical assay feasibility was confirmed through genital secretion samples from patients suspected of N. gonorrhoeae infection, and the outcomes were compared to the N. gonorrhoeae-qPCR technique. Our assay's sensitivity and specificity were 100% (95% CI: 93.28 to 100.00%) and 100% (95% CI: 81.47 to 100.00%). Taken together, the preliminary results indicated that our PfAgo-N. gonorrhoeae assay can act as a practical point-of-care (POC) diagnostic method, enabling accurate and convenient detection of N. gonorrhoeae infection, particularly in resource-limited settings.