Abstract: Bacterial infections pose a major challenge in the field of public health, making the development of rapid and accurate detection technologies crucial for disease prevention and control. Conventional methods such as culture, PCR (polymerase chain reaction), and ELISA (enzyme-linked immunosorbent assay) are limited by long processing times, limited sensitivity, and the inability to perform real-time monitoring. In contrast, fluorescent probe technology, with its high specificity and sensitivity, enables rapid and visual detection of bacteria by recognizing bacterial targets and generating fluorescence signals. This review systematically summarizes the molecular design strategies, luminescence mechanisms, and classifications of fluorescent probes, with a focus on their current applications in medical diagnosis, food safety, and environmental monitoring. It also highlights the challenges the technology faces, including detection limits, specificity, and clinical translation. Looking ahead, innovative approaches such as multimodal probe integration and artificial intelligence-assisted analysis are expected to drive the technology toward more intelligent and practical advancements. Fluorescent probe technology provides a powerful tool for research and control of bacterial-related issues.