Decoding biochemical fingerprints: Artificial intelligence-empowered Raman spectroscopy for rapid phenotypic...

Rapid phenotypic antimicrobial susceptibility testing (AST) is urgently needed to combat the escalating crisis of antimicrobial resistance (AMR). Raman spectroscopy, a label-free vibrational technique, captures phenotypic "biochemical fingerprints" of pathogens but is hindered in complex clinical matrices by low signal-to-noise ratio (SNR), strong background fluorescence, and batch-to-batch variation. To overcome these analytical bottlenecks, artificial intelligence (AI), particularly deep learning, has emerged as a transformative computational paradigm. This review proposes a "hardware-algorithm synergistic" evaluation framework for AI-assisted Raman AST. First, we systematically assess physical trade-offs among Raman platforms (RS, SERS, and LTRS) and analyze feature-learning mechanisms of AI architectures, including deep convolutional networks, Transformers, and domain-adaptive transfer learning. Notably, Transformers present a critical trade-off between capturing long-range dependencies and computational overhead for POCT. Then, we discuss explainable AI that maps mathematical decisions to underlying biochemical mechanisms, along with closed-loop diagnostic systems integrating microfluidics and Raman-activated cell sorting (RACS). Finally, we highlight future directions, such as Raman foundation models and multi-modal cross-scale fusion, aiming to provide a theoretical roadmap to dismantle translational barriers of rapid AST in clinical practice.