<p>Surface plasmon resonance (SPR) biosensoris a new type of high sensitivity, real-time, label-free detection technology, which plays an increasingly important role in the biomedicine field. Considering the urgent requirement of trace detection, especially for diagnosing early-stage diseases, the demand for high detection sensitivity of sensors is increasing. In recent years, various nanostructures have been proposed to design SPR biosensors. By constructing composite nanostructures, the detection sensitivity has been significantly enhanced, which has become a promising solution to expand the application of SPR biosensors. This review systematically summarized the basic principle, fabrication and illustrative application of SPR biosensors based on versatile nanostructures. Firstly, the mechanisms of various nanostructures to enhance the detection sensitivity of SPR biosensors were clarified. Then, the preparation strategies of various nanostructures were comprehensively illustrated. In addition, this review also summarized the latest applications of SPR biosensors with different structures. Finally, this review carefully highlighted the current challenges and possible development directions in future.</p>