The evolution of next-generation lateral flow assays for bacterial and fungal diagnostics

Infectious diseases driven by increasingly resistant bacterial and fungal pathogens demand diagnostics that are faster and more accessible than conventional culture-based methods. This review traces the evolution of lateral flow assays (LFAs) from simple qualitative strips to sophisticated, molecularly enhanced diagnostic platforms. This review synthesizes literature mostly published between 2010 and 2025, identified through PubMed, Scopus, and Web of Science using search terms including 'lateral flow assay,' 'point-of-care diagnostics,' 'CRISPR diagnostics,' 'nanozyme biosensor,' 'antimicrobial resistance,' 'Candida auris,' and 'invasive aspergillosis. It highlights how advances in materials science (including quantum dots and nanozymes), isothermal amplification (RPA, LAMP), and CRISPR/Cas-based recognition have pushed LFAs toward laboratory-comparable sensitivity while preserving their simplicity. The clinical impact of these next-generation LFAs is illustrated using high-threat pathogens such as MRSA, Candida auris, and invasive Aspergillus, where rapid, point-of-care identification improves outcomes and supports antimicrobial stewardship. The review also examines the digital transformation of LFAs through smartphone-based readouts and artificial intelligence, which enable quantitative analysis and real-time epidemiological surveillance, even in remote settings. Despite ongoing challenges, including the hook effect, cross-reactivity, and regulatory fragmentation, the emerging technologies described here suggest that LFAs can help decouple high-quality infectious disease diagnostics from centralized laboratories, supporting a more equitable, global access to precision microbiological testing.