Fluorescence spectroscopy is an important testing method. Recently, the optical-fiber-probe-based fluorescence testing technology has become a popular research topic. This technology exhibits the advantages of high efficiency, micro, real-time, in-situ, small size, and easy integration. This review briefly describes the principle of fluorescence analysis, the spatial conduction theory of laser emission and fluorescence collection using optical-fiber probes, the typical structure and preparation of the optical-fiber fluorescent probes, and the status of research in the fields of biology, environment, and food safety. Finally, this review looks forward to the development trend of optical-fiber fluorescent probes.