Exploition of novel antibacterial agents that contact non-specifically with bacteria is one of the solutions to the problem of bacterial infection. Firstly, guanidine-based antibacterial polymers having prolonged, broad-spectrum, high-efficiency antibacterial properties, without eukaryotic cytotoxicity, and making it difficult for bacteria to develop resistance are introduced briefly. Then, the antibacterial mechanism of non-specific electrostatic attraction with bacteria is reviewed. In addition, the design, synthesis, and antimicrobial property of the main-chained guanidine-based antibacterial polymers, side-chained guanidine-based antibacterial polymers, and surface-grafted guanidine-based antibacterial polymers are described in detail. Finally, the future development of controllable synthetic strategy and practical application of the novel guanidine-based antibacterial polymers are prospected.