Metal-enhanced fluorescence (MEF) is a unique phenomenon of surface plasmons, where light interacts with the metallic nanostructures and produces electromagnetic fields to enhance the sensitivity of fluorescence-based detection. In particular, this enhancement in sensing capacity is of importance to many research areas, including medical diagnostics, forensic science, and biotechnology. The article covers the basic mechanism of MEF and recent developments in plasmonic nanostructures fabrication for efficient fluorescence signal enhancement that are critically reviewed. The implications of current fluorescence-based technologies for biosensors are summarized, which are in practice to detect different analytes relevant to food control, medical diagnostics, and forensic science. Furthermore, characteristics of existing fabrication methods have been compared on the basis of their resolution, design flexibility, and throughput. The future projections emphasize exploring the potential of non-conventional materials and hybrid fabrication techniques to further enhance the sensitivity of MEF-based biosensors.